Oral delivery of modified transferrin fusion proteins

ABSTRACT

Pharmaceutical compositions containing modified fusion proteins of transferrin and therapeutic proteins or peptides with increased serum half-life or increased serum stability are disclosed. Preferred fusion proteins include those modified so that the transferrin moiety exhibits no or reduced glycosylation, but does exhibit binding to iron and/or the transferrin receptor. Such fusion proteins may be administered orally.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application60/460,829, filed on Apr. 8, 2003, U.S. application Ser. No. 10/378,094,filed Mar. 4, 2003, and U.S. Provisional Application 60/406,977, filedAug. 30, 2002, all of which are herein incorporated by reference intheir entirety.

FIELD OF THE INVENTION

The present invention relates to orally administerable therapeuticproteins or peptides with extended serum stability or serum half-life,particularly to therapeutic proteins or peptides fused to or inserted ina transferrin molecule modified to reduce or inhibit glycosylation.

BACKGROUND OF THE INVENTION

Therapeutic proteins or peptides in their native state or whenrecombinantly produced are typically labile molecules exhibiting shortperiods of serum stability or short in vivo circulatory half-lives. Inaddition, these molecules are often extremely labile when formulated,particularly when formulated in aqueous solutions for diagnostic andtherapeutic purposes.

Few practical solutions exist to extend or promote the stability in vivoor in vitro of proteinaceous therapeutic molecules. Polyethylene glycol(PEG) is a substance that can be attached to a protein, resulting inlonger-acting, sustained activity of the protein. If the activity of aprotein is prolonged by the attachment to PEG, the frequency that theprotein needs to be administered may be decreased. PEG attachment,however, often decreases or destroys the protein's therapeutic activity.While in some instance PEG attachment can reduce immunogenicity of theprotein, in other instances it may increase immunogenicity.

Therapeutic proteins or peptides have also been stabilized by fusion toa protein capable of extending the in vivo circulatory half-life of thetherapeutic protein. For instance, therapeutic proteins fused to albuminor to antibody fragments may exhibit extended in vivo circulatoryhalf-life when compared to the therapeutic protein in the unfused state.See U.S. Pat. Nos. 5,876,969 and 5,766,883.

Another serum protein, glycosylated human transferrin (Tf) has also beenused to make fusions with therapeutic proteins to target delivery to theinterior of cells or to carry agents across the blood-brain barrier.These fusion proteins comprising glycosylated human Tf have been used totarget nerve growth factor (NGF) or ciliary neurotrophic factor (CNTF)across the blood-brain barrier by fusing full-length Tf to the agent.See U.S. Pat. Nos. 5,672,683 and 5,977,307. In these fusion proteins,the Tf portion of the molecule is glycosylated and binds to two atoms ofiron, which is required for Tf binding to its receptor on a cell and,according to the inventors of these patents, to target delivery of theNGF or CNTF moiety across the blood-brain barrier. Transferrin fusionproteins have also been produced by inserting an HIV-1 protease targetsequence into surface exposed loops of glycosylated transferrin toinvestigate the ability to produce another form of Tf fusion fortargeted delivery to the inside of a cell via the Tf receptor (Ali etal. (1999) J. Biol. Chem. 274(34):24066-24073).

Serum transferrin (Tf) is a monomeric glycoprotein with a molecularweight of 80,000 daltons that binds iron in the circulation andtransports it to various tissues via the transferrin receptor (TfR)(Aisen et al. (1980) Ann. Rev. Biochem. 49: 357-393; MacGillivray et al.(1981) J. Biol. Chem. 258: 3543-3553, U.S. Pat. No. 5,026,651). Tf isone of the most common serum molecules, comprising up to about 5-10% oftotal serum proteins. Carbohydrate deficient transferrin occurs inelevated levels in the blood of alcoholic individuals and exhibits alonger half life (approximately 14-17 days) than that of glycosylatedtransferrin (approximately 7-10 days). See van Eijk et al. (1983) Clin.Chim. Acta 132:167-171, Stibler (1991) Clin. Chem. 37:2029-2037 (1991),Arndt (2001) Clin. Chem. 47(1):13-27 and Stibler et al. in“Carbohydrate-deficient consumption”, Advances in the Biosciences, (EdNordmann et al.), Pergamon, 1988, Vol. 71, pages 353-357).

The structure of Tf has been well characterized and the mechanism ofreceptor binding, iron binding and release and carbonate ion bindinghave been elucidated (U.S. Pat. Nos. 5,026,651, 5,986,067 andMacGillivray et al. (1983) J. Biol. Chem. 258(6):3543-3546).

Transferrin and antibodies that bind the transferrin receptor have alsobeen used to deliver or carry toxic agents to tumor cells as cancertherapy (Baselga and Mendelsohn, 1994), and transferrin has been used asa non-viral gene therapy vector to deliver DNA to cells (Frank et al.,1994; Wagner et al., 1992). The ability to deliver proteins to thecentral nervous system (CNS) using the transferrin receptor as the entrypoint has been demonstrated with several proteins and peptides includingCD4 (Walus et al., 1996), brain derived neurotrophic factor (Pardridgeet al., 1994), glial derived neurotrophic factor (Albeck et al.), avasointestinal peptide analogue (Bickel et al., 1993), a beta-amyloidpeptide (Saito et al., 1995), and an antisense oligonucleotide(Pardridge et al., 1995).

Transferrin fusion proteins have not, however, been modified orengineered to extend the in vivo circulatory half-life of a therapeuticprotein nor peptide or to increase bioavailability by reducing orinhibiting glycosylation of the Tf moiety nor to reduce or prevent ironand/or Tf receptor binding.

SUMMARY OF THE INVENTION

As described in more detail below, the present invention includes orallyadministerable modified Tf fusion proteins comprising at least onetherapeutic protein, polypeptide or peptide entity, wherein the Tfportion is engineered to extend the serum half-life or bioavailabilityof the molecule. The invention also includes pharmaceutical formulationsand compositions formulated for oral administration comprising thefusion proteins, methods of extending the serum stability, serumhalf-life and bioavailability of a therapeutic protein by fusion tomodified transferrin, nucleic acid molecules encoding the modified Tffusion proteins, and the like. Moreover, the present invention relatesto methods of treating a patient with a modified Tf fusion protein byoral administration. Further, the present invention relates to methodsof treating a patient with a modified Tf fusion protein by intranasaladministration. Additionally, the present invention relates to methodsof treating a patient with a modified Tf fusion protein by pulmonaryadministration.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an alignment of the N and C Domains of Human (Hu)transferrin (Tf) (SEQ ID NO: 3) with similarities and identitieshighlighted.

FIG. 2A-2B shows an alignment of transferrin sequences from differentspecies (SEQ ID NOs: 3 and 48-54). Light shading: Similarity; Darkshading: Identity.

FIG. 3 shows the location of a number of Tf surface exposed insertionsites for therapeutic proteins, polypeptides or peptides.

DETAILED DESCRIPTION General Description

It has been discovered that a therapeutic protein (e.g., a polypeptide,antibody, or peptide, or fragments and variants thereof) can bestabilized to extend the in vivo circulatory half-life and/or retain thetherapeutic protein's activity for extended periods of time in vivo bygenetically fusing or chemically conjugating the therapeutic protein,polypeptide or peptide to all or a portion of modified transferrinsufficient to extend its half life in serum. The modified transferrinfusion proteins include a transferrin protein or domain covalentlylinked to a therapeutic protein or peptide, wherein the transferrinportion is modified to contain one or more amino acid substitutions,insertions or deletions compared to a wild-type transferrin sequence. Inone embodiment, Tf fusion proteins are engineered to reduce or preventglycosylation within the Tf or a Tf domain. In other embodiments, the Tfprotein or Tf domain(s) is modified to exhibit reduced or no binding toiron or carbonate ion, or to have a reduced affinity or not bind to a Tfreceptor (TfR).

The present invention therefore includes transferrin fusion proteins,therapeutic compositions comprising the fusion proteins, and methods oftreating, preventing, or ameliorating diseases or disorders byadministering the fusion proteins. A transferrin fusion protein of theinvention includes at least a fragment or variant of a therapeuticprotein and at least a fragment or variant of modified transferrin,which are associated with one another, preferably by genetic fusion(i.e., the transferrin fusion protein is generated by translation of anucleic acid in which a polynucleotide encoding all or a portion of atherapeutic protein is joined in-frame with a polynucleotide encodingall or a portion of modified transferrin) or chemical conjugation to oneanother. The therapeutic protein and transferrin protein, once part ofthe transferrin fusion protein, may be referred to as a “portion”,“region” or “moiety” of the transferrin fusion protein (e.g., a“therapeutic protein portion’ or a “transferrin protein portion”).

In one embodiment, the invention provides a transferrin fusion proteincomprising, or alternatively consisting of, a therapeutic protein and amodified serum transferrin protein. In other embodiments, the inventionprovides a transferrin fusion protein comprising, or alternativelyconsisting of, a biologically active and/or therapeutically activefragment of a therapeutic protein and a modified transferrin protein. Inother embodiments, the invention provides a transferrin fusion proteincomprising, or alternatively consisting of, a biologically active and/ortherapeutically active variant of a therapeutic protein and modifiedtransferrin protein. In further embodiments, the invention provides atransferrin fusion protein comprising a therapeutic protein, and abiologically active and/or therapeutically active fragment of modifiedtransferrin. In another embodiment, the therapeutic protein portion ofthe transferrin fusion protein is the active form of the therapeuticprotein.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are described.

DEFINITIONS

As used herein, an “amino acid corresponding to” or an “equivalent aminoacid” in a transferrin sequence is identified by alignment to maximizethe identity or similarity between a first transferrin sequence and atleast a second transferrin sequence. The number used to identify anequivalent amino acid in a second transferrin sequence is based on thenumber used to identify the corresponding amino acid in the firsttransferrin sequence. In certain cases, these phrases may be used todescribe the amino acid residues in human transferrin compared tocertain residues in rabbit serum transferrin.

As used herein, the term “biological activity” refers to a function orset of activities performed by a therapeutic molecule, protein orpeptide in a biological context (i.e., in an organism or an in vitrofacsimile thereof). Biological activities may include but are notlimited to the functions of the therapeutic molecule portion of theclaimed fusion proteins, such as, but not limited to, the induction ofextracellular matrix secretion from responsive cell lines, the inductionof hormone secretion, the induction of chemotaxis, the induction ofmitogenesis, the induction of differentiation, or the inhibition of celldivision of responsive cells. A fusion protein or peptide of theinvention is considered to be biologically active if it exhibits one ormore biological activities of its therapeutic protein's nativecounterpart.

As used herein, “binders” are agents used to impart cohesive qualitiesto the powdered material. Binders, or “granulators” as they aresometimes known, impart a cohesiveness to the tablet formulation, whichinsures the tablet remaining intact after compression, as well asimproving the free-flowing qualities by the formulation of granules ofdesired hardness and size. Materials commonly used as binders includestarch; gelatin; sugars, such as sucrose, glucose, dextrose, molasses,and lactose; natural and synthetic gums, such as acacia, sodiumalginate, extract of Irish moss, panwar gum, ghatti gum, mucilage ofisapol husks, carboxymethylcellulose, methylcellulose,polyvinylpyrrolidone, Veegum, microcrystalline cellulose,microcrystalline dextrose, amylose, and larch arabogalactan, and thelike.

As used herein, the term “carrier” refers to a diluent, adjuvant,excipient, or vehicle with which a composition is administered. Suchpharmaceutical carriers can be sterile liquids, such as water and oils,including those of petroleum, animal, vegetable or synthetic origin,such as peanut oil, soybean oil, mineral oil, sesame oil and the like.

As used herein, “coloring agents” are agents that give tablets a morepleasing appearance, and in addition help the manufacturer to controlthe product during its preparation and help the user to identify theproduct. Any of the approved certified water-soluble FD&C dyes, mixturesthereof, or their corresponding lakes may be used to color tablets. Acolor lake is the combination by adsorption of a water-soluble dye to ahydrous oxide of a heavy metal, resulting in an insoluble form of thedye.

As used herein, “diluents” are inert substances added to increase thebulk of the formulation to make the tablet a practical size forcompression. Commonly used diluents include calcium phosphate, calciumsulfate, lactose, kaolin, mannitol, sodium chloride, dry starch,powdered sugar, silica, and the like.

As used herein, “disintegrators” or “disintegrants” are substances thatfacilitate the breakup or disintegration of tablets afteradministration. Materials serving as disintegrants have been chemicallyclassified as starches, clays, celluloses, algins, or gums. Otherdisintegrators include Veegum HV, methylcellulose, agar, bentonite,cellulose and wood products, natural sponge, cation-exchange resins,alginic acid, guar gum, citrus pulp, cross-linked polyvinylpyrrolidone,carboxymethylcellulose, and the like.

As used herein, the term “dispersibility” or “dispersible” means a drypowder having a moisture content of less than about 10% by weight (% w)water, usually below about 5% w and preferably less than about 3% w; aparticle size of about 1.0-5.0 μm mass median diameter (MMD), usually1.0-4.0 μm MMD, and preferably 1.0-3.0 μm MMD; a delivered dose ofabout >30%, usually >40%, preferably >50%, and most preferred >60%; andan aerosol particle size distribution of 1.0-5.0 μm mass medianaerodynamic diameter (MMAD), usually 1.5-4.5 μm MMAD, and preferably1.5-4.0 μm MMAD.

As used herein, the term “dry” means that the composition has a moisturecontent such that the particles are readily dispersible in an inhalationdevice to form an aerosol. This moisture content is generally belowabout 10% by weight (% w) water, usually below about 5% w and preferablyless than about 3% w.

As used herein, “effective amount” means an amount of a drug orpharmacologically active agent that is sufficient to provide the desiredlocal or systemic effect and performance at a reasonable benefit/riskratio attending any medical treatment.

As used herein, “flavoring agents” vary considerably in their chemicalstructure, ranging from simple esters, alcohols, and aldehydes tocarbohydrates and complex volatile oils. Synthetic flavors of almost anydesired type are now available.

As used herein, the terms “fragment of a Tf protein” or “Tf protein,” or“portion of a Tf protein” refer to an amino acid sequence comprising atleast about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%,97%, 98%, 99% or 100% of a naturally occurring Tf protein or mutantthereof.

As used herein, the term “gene” refers to any segment of DNA associatedwith a biological function. Thus, genes include, but are not limited to,coding sequences and/or the regulatory sequences required for theirexpression. Genes can also include non-expressed DNA segments that, forexample, form recognition sequences for other proteins. Genes can beobtained from a variety of sources, including cloning from a source ofinterest or synthesizing from known or predicted sequence information,and may include sequences designed to have desired parameters.

As used herein, a “heterologous polynucleotide” or a “heterologousnucleic acid” or a “heterologous gene” or a “heterologous sequence” oran “exogenous DNA segment” refers to a polynucleotide, nucleic acid orDNA segment that originates from a source foreign to the particular hostcell, or, if from the same source, is modified from its original form. Aheterologous gene in a host cell includes a gene that is endogenous tothe particular host cell, but has been modified. Thus, the terms referto a DNA segment which is foreign or heterologous to the cell, orhomologous to the cell but in a position within the host cell nucleicacid in which the element is not ordinarily found. As an example, asignal sequence native to a yeast cell but attached to a human Tfsequence is heterologous.

As used herein, an “isolated” nucleic acid sequence refers to a nucleicacid sequence which is essentially free of other nucleic acid sequences,e.g., at least about 20% pure, preferably at least about 40% pure, morepreferably about 60% pure, even more preferably about 80% pure, mostpreferably about 90% pure, and even most preferably about 95% pure, asdetermined by agarose gel electrophoresis. For example, an isolatednucleic acid sequence can be obtained by standard cloning proceduresused in genetic engineering to relocate the nucleic acid sequence fromits natural location to a different site where it will be reproduced.The cloning procedures may involve excision and isolation of a desirednucleic acid fragment comprising the nucleic acid sequence encoding thepolypeptide, insertion of the fragment into a vector molecule, andincorporation of the recombinant vector into a host cell where multiplecopies or clones of the nucleic acid sequence will be replicated. Thenucleic acid sequence may be of genomic, cDNA, RNA, semi-synthetic,synthetic origin, or any combinations thereof.

As used herein, two or more DNA coding sequences are said to be “joined”or “fused” when, as a result of in-frame fusions between the DNA codingsequences, the DNA coding sequences are translated into a fusionpolypeptide. The term “fusion” in reference to Tf fusions includes, butis not limited to, attachment of at least one therapeutic protein,polypeptide or peptide to the N-terminal end of Tf, attachment to theC-terminal end of Tf, and/or insertion between any two amino acidswithin Tf.

As used herein, “lubricants” are materials that perform a number offunctions in tablet manufacture, such as improving the rate of flow ofthe tablet granulation, preventing adhesion of the tablet material tothe surface of the dies and punches, reducing interparticle friction,and facilitating the ejection of the tablets from the die cavity.Commonly used lubricants include talc, magnesium stearate, calciumstearate, stearic acid, and hydrogenated vegetable oils. Typical amountsof lubricants range from about 0.1% by weight to about 5% by weight.

As used herein, “Modified transferrin” as used herein refers to atransferrin molecule that exhibits at least one modification of itsamino acid sequence, compared to wild-type transferrin.

As used herein, “Modified transferrin fusion protein” as used hereinrefers to a protein formed by the fusion of at least one molecule ofmodified transferrin (or a fragment or variant thereof) to at least onemolecule of a therapeutic protein (or fragment or variant thereof).

As used herein, the terms “nucleic acid” or “polynucleotide” refer todeoxyribonucleotides or ribonucleotides and polymers thereof in eithersingle- or double-stranded form. Unless specifically limited, the termsencompass nucleic acids containing analogues of natural nucleotides thathave similar binding properties as the reference nucleic acid and aremetabolized in a manner similar to naturally occurring nucleotides.Unless otherwise indicated, a particular nucleic acid sequence alsoimplicitly encompasses conservatively modified variants thereof (e.g.degenerate codon substitutions) and complementary sequences as well asthe sequence explicitly indicated. Specifically, degenerate codonsubstitutions may be achieved by generating sequences in which the thirdposition of one or more selected (or all) codons is substituted withmixed-base and/or deoxyinosine residues (Batzer et al. (1991) NucleicAcid Res. 19:5081; Ohtsuka et al. (1985) J. Biol. Chem. 260:2605-2608;Cassol et al. (1992); Rossolini et al. (1994) Mol. Cell. Probes8:91-98). The term nucleic acid is used interchangeably with gene, cDNA,and mRNA encoded by a gene.

As used herein, a DNA segment is referred to as “operably linked” whenit is placed into a functional relationship with another DNA segment.For example, DNA for a signal sequence is operably linked to DNAencoding a fusion protein of the invention if it is expressed as apreprotein that participates in the secretion of the fusion protein; apromoter or enhancer is operably linked to a coding sequence if itstimulates the transcription of the sequence. Generally, DNA sequencesthat are operably linked are contiguous, and in the case of a signalsequence or fusion protein both contiguous and in reading phase.However, enhancers need not be contiguous with the coding sequenceswhose transcription they control. Linking, in this context, isaccomplished by ligation at convenient restriction sites or at adaptersor linkers inserted in lieu thereof.

As used herein, “pharmaceutically acceptable” refers to materials andcompositions that are physiologically tolerable and do not typicallyproduce an allergic or similar untoward reaction, such as gastric upset,dizziness and the like, when administered to a human. Typically, as usedherein, the term “pharmaceutically acceptable” means approved by aregulatory agency of the Federal or a state government or listed in theU.S. Pharmacopeia or other generally recognized pharmacopeia for use inanimals, and more particularly in humans.

As used herein, “physiologically effective amount” is that amountdelivered to a subject to give the desired palliative or curativeeffect. This amount is specific for each drug and its ultimate approveddosage level.

As used herein, the term “powder” means a composition that consists offinely dispersed solid particles that are free flowing and capable ofbeing readily dispersed in an inhalation device and subsequently inhaledby a subject so that the particles reach the lungs to permit penetrationinto the alveoli. Thus, the powder is said to be “respirable.”Preferably the average particle size is less than about 10 microns (μm)in diameter with a relatively uniform spheroidal shape distribution.More preferably the diameter is less than about 7.5 μm and mostpreferably less than about 5.0 μm. Usually the particle sizedistribution is between about 0.1 μm and about 5 μm in diameter,particularly about 0.3 μm to about 5 μm.

As used herein, the term “promoter” refers to a region of DNA involvedin binding RNA polymerase to initiate transcription.

As used herein, the term “recombinant” refers to a cell, tissue ororganism that has undergone transformation with a new combination ofgenes or DNA.

As used herein, the term “subject” can be a human, a mammal, or ananimal. The subject being treated is a patient in need of treatment.

As used herein, a targeting entity, protein, polypeptide or peptiderefers to a molecule that binds specifically to a particular cell type[normal (e.g., lymphocytes) or abnormal e.g., (cancer cell)] andtherefore may be used to target a Tf fusion protein or compound (drug,or cytotoxic agent) to that cell type specifically.

As used herein, “tablets” are solid pharmaceutical dosage formscontaining drug substances with or without suitable diluents andprepared either by compression or molding methods well known in the art.Tablets have been in widespread use since the latter part of the 19^(th)century and their popularity continues. Tablets remain popular as adosage form because of the advantages afforded both to the manufacturer(e.g., simplicity and economy of preparation, stability; and conveniencein packaging, shipping, and dispensing) and the patient (e.g., accuracyof dosage, compactness, portability, blandness of taste, and ease ofadministration). Although tablets are most frequently discoid in shape,they may also be round, oval, oblong, cylindrical, or triangular. Theymay differ greatly in size and weight depending on the amount of drugsubstance present and the intended method of administration. They aredivided into two general classes, (1) compressed tablets, and (2) moldedtablets or tablet triturates. In addition to the active or therapeuticingredient or ingredients, tablets contain a number or inert materialsor additives. A first group of such additives includes those materialsthat help to impart satisfactory compression characteristics to theformulation, including diluents, binders, and lubricants. A second groupof such additives helps to give additional desirable physicalcharacteristics to the finished tablet, such as disintegrators, colors,flavors, and sweetening agents.

As used herein, the term “therapeutically effective amount” refers tothat amount of the transferrin fusion protein comprising a therapeuticmolecule which, when administered to a subject in need thereof, issufficient to effect treatment. The amount of transferrin fusion proteinwhich constitutes a “therapeutically effective amount” will varydepending on the therapeutic protein used, the severity of the conditionor disease, and the age and body weight of the subject to be treated,but can be determined routinely by one or ordinary skill in the arthaving regard to his/her own knowledge and to this disclosure.

As used herein, “therapeutic protein” refers to proteins, polypeptides,peptides or fragments or variants thereof, having one or moretherapeutic and/or biological activities. Therapeutic proteinsencompassed by the invention include but are not limited to proteins,polypeptides, peptides, antibodies, and biologics. The terms peptides,proteins, and polypeptides are used interchangeably herein.Additionally, the term “therapeutic protein” may refer to the endogenousor naturally occurring correlate of a therapeutic protein. By apolypeptide displaying a “therapeutic activity” or a protein that is“therapeutically active” is meant a polypeptide that possesses one ormore known biological and/or therapeutic activities associated with atherapeutic protein such as one or more of the therapeutic proteinsdescribed herein or otherwise known in the art. As a non-limitingexample, a “therapeutic protein” is a protein that is useful to treat,prevent or ameliorate a disease, condition or disorder. Such a disease,condition or disorder may be in humans or in a non-human animal, e.g.,veterinary use.

As used herein, the term “transformation” refers to the transfer ofnucleic acid (i.e., a nucleotide polymer) into a cell. As used herein,the term “genetic transformation” refers to the transfer andincorporation of DNA, especially recombinant DNA, into a cell. As usedherein, the term “transformant” refers to a cell, tissue or organismthat has undergone transformation.

As used herein, the term “transgene” refers to a nucleic acid that isinserted into an organism, host cell or vector in a manner that ensuresits function. As used herein, the term “transgenic” refers to cells,cell cultures, organisms, bacteria, fungi, animals, plants, and progenyof any of the preceding, which have received a foreign or modified geneand in particular a gene encoding a modified Tf fusion protein by one ofthe various methods of transformation, wherein the foreign or modifiedgene is from the same or different species than the species of theorganism receiving the foreign or modified gene.

As used herein, the term “Variants or variant” refers to apolynucleotide or nucleic acid differing from a reference nucleic acidor polypeptide, but retaining essential properties thereof. Generally,variants are overall closely similar, and, in many regions, identical tothe reference nucleic acid or polypeptide. As used herein, “variant”refers to a therapeutic protein portion of a transferrin fusion proteinof the invention, differing in sequence from a native therapeuticprotein but retaining at least one functional and/or therapeuticproperty thereof as described elsewhere herein or otherwise known in theart.

As used herein, the term “vector” refers broadly to any plasmid,phagemid or virus encoding an exogenous nucleic acid. The term is alsobe construed to include non-plasmid, non-phagemid and non-viralcompounds which facilitate the transfer of nucleic acid into virions orcells, such as, for example, polylysine compounds and the like. Thevector may be a viral vector that is suitable as a delivery vehicle fordelivery of the nucleic acid, or mutant thereof, to a cell, or thevector may be a non-viral vector which is suitable for the same purpose.Examples of viral and non-viral vectors for delivery of DNA to cells andtissues are well known in the art and are described, for example, in Maet al. (1997, Proc. Natl. Acad. Sci. U.S.A. 94:12744-12746). Examples ofviral vectors include, but are not limited to, a recombinant vacciniavirus, a recombinant adenovirus, a recombinant retrovirus, a recombinantadeno-associated virus, a recombinant avian pox virus, and the like(Cranage et al., 1986, EMBO 3. 5:3057-3063; International PatentApplication No. WO94/17810, published Aug. 18, 1994; InternationalPatent Application No. WO94/23744, published Oct. 27, 1994). Examples ofnon-viral vectors include, but are not limited to, liposomes, polyaminederivatives of DNA, and the like.

As used herein, the term “wild type” refers to a polynucleotide orpolypeptide sequence that is naturally occurring.

Transferrin and Transferrin Modifications

Any transferrin may be used to make modified Tf fusion proteins of theinvention. As an example, the wild-type human Tf (Tf) is a 679 aminoacid protein of approximately 75 kDa (not accounting for glycosylation),with two main domains, N (about 330 amino acids) and C (about 340 aminoacids), which appear to originate from a gene duplication. See GenBankaccession numbers NM001063, XM002793, M12530, XM039845, XM 039847 andS95936 (www.ncbi.nlm.nih.gov/), all of which are herein incorporated byreference in their entirety, as well as SEQ ID NOS 1, 2 and 3. The twodomains have diverged over time but retain a large degree ofidentity/similarity (FIG. 1).

Each of the N and C domains is further divided into two subdomains, N1and N2, C1 and C2. The function of Tf is to transport iron to the cellsof the body. This process is mediated by the Tf receptor (TfR), which isexpressed on all cells, particularly actively growing cells. TfRrecognizes the iron bound form of Tf (two molecules of which are boundper receptor), endocytosis then occurs whereby the TfR/Tf complex istransported to the endosome, at which point the localized drop in pHresults in release of bound iron and the recycling of the TfR/Tf complexto the cell surface and release of Tf (known as apoTf in its un-ironbound form). Receptor binding is through the C domain of Tf. The twoglycosylation sites in the C domain do not appear to be involved inreceptor binding as unglycosylated iron bound Tf does bind the receptor.

Each Tf molecule can carry two iron ions (Fe³⁺). These are complexed inthe space between the N1 and N2, C1 and C2 sub domains resulting in aconformational change in the molecule. Tf crosses the blood brainbarrier (BBB) via the Tf receptor.

In human transferrin, the iron binding sites comprise at least aminoacids Asp 63 (Asp 82 of SEQ ID NO: 2 which includes the native Tf signalsequence), Asp 392 (Asp 411 of SEQ ID NO: 2), Tyr 95 (Tyr 114 of SEQ IDNO: 2), Tyr 426 (Tyr 445 of SEQ ID NO: 2), Tyr 188 (Tyr 207 of SEQ IDNO: 2), Tyr 514 or 517 (Tyr 533 or Tyr 536 SEQ ID NO: 2), His 249 (His268 of SEQ ID NO: 2), and His 585 (His 604 of SEQ ID NO: 2) of SEQ IDNO: 3. The hinge regions comprise at least N domain amino acid residues94-96, 245-247 and/or 316-318 as well as C domain amino acid residues425-427, 581-582 and/or 652-658 of SEQ ID NO: 3. The carbonate bindingsites comprise at least amino acids Thr 120 (Thr 139 of SEQ ID NO: 2),Thr 452 (Thr 471 of SEQ ID NO: 2), Arg 124 (Arg 143 of SEQ ID NO: 2),Arg 456 (Arg 475 of SEQ ID NO: 2), Ala 126 (Ala 145 of SEQ ID NO: 2),Ala 458 (Ala 477 of SEQ ID NO: 2), Gly 127 (Gly 146 of SEQ ID NO: 2),and Gly 459 (Gly 478 of SEQ ID NO: 2) of SEQ ID NO: 3.

In one embodiment of the invention, the modified transferrin fusionprotein includes a modified human transferrin, although any animal Tfmolecule may be used to produce the fusion proteins of the invention,including human Tf variants, cow, pig, sheep, dog, rabbit, rat, mouse,hamster, echnida, platypus, chicken, frog, hornworm, monkey, as well asother bovine, canine and avian species. All of these Tf sequences arereadily available in GenBank and other public databases. The human Tfnucleotide sequence is available (see SEQ ID NOS 1, 2 and 3 and theaccession numbers described above and available atwww.ncbi.nlm.nih.gov/) and can be used to make genetic fusions betweenTf or a domain of Tf and the therapeutic molecule of choice. Fusions mayalso be made from related molecules such as lacto transferrin(lactoferrin) GenBank Acc: NM_(—)002343) or melanotransferrin (GenBankAcc. NM_(—)013900, murine melanotransferrin).

Melanotransferrin is a glycosylated protein found at high levels inmalignant melanoma cells and was originally named human melanoma antigenp97 (Brown et al., 1982, Nature, 296: 171-173). It possesses highsequence homology with human serum transferrin, human lactoferrin, andchicken transferrin (Brown et al., 1982, Nature, 296: 171-173; Rose etal., Proc. Natl. Acad. Sci. USA, 1986, 83: 1261-1265). However, unlikethese receptors, no cellular receptor has been identified formelanotransferrin. Melanotransferrin reversibly binds iron and it existsin two forms, one of which is bound to cell membranes by a glycosylphosphatidylinositol anchor while the other form is both soluble andactively secreted (Baker et al., 1992, FEBS Lett, 298: 215-218; Alemanyet al., 1993, J. Cell Sci., 104: 1155-1162; Food et al., 1994, J. Biol.Chem. 274: 7011-7017).

Lactoferrin (Lf), a natural defense iron-binding protein, has been foundto possess antibacterial, antimycotic, antiviral, antineoplastic andanti-inflammatory activity. The protein is present in exocrinesecretions that are commonly exposed to normal flora: milk, tears, nasalexudate, saliva, bronchial mucus, gastrointestinal fluids,cervico-vaginal mucus and seminal fluid. Additionally, Lf is a majorconstituent of the secondary specific granules of circulatingpolymorphonuclear neutrophils (PMNs). The apoprotein is released ondegranulation of the PMNs in septic areas. A principal function of Lf isthat of scavenging free iron in fluids and inflamed areas so as tosuppress free radical-mediated damage and decrease the availability ofthe metal to invading microbial and neoplastic cells. In a study thatexamined the turnover rate of ¹²⁵I Lf in adults, it was shown that Lf israpidly taken up by the liver and spleen, and the radioactivitypersisted for several weeks in the liver and spleen (Bennett et al.(1979), Clin. Sci. (Lond.) 57: 453-460).

In one embodiment, the transferrin portion of the transferrin fusionprotein of the invention includes a transferrin splice variant. In oneexample, a transferrin splice variant can be a splice variant of humantransferrin. In one specific embodiment, the human transferrin splicevariant can be that of Genbank Accession AAA61140.

In another embodiment, the transferrin portion of the transferrin fusionprotein of the invention includes a lactoferrin splice variant. In oneexample, a human serum lactoferrin splice variant can be a novel splicevariant of a neutrophil lactoferrin. In one specific embodiment, theneutrophil lactoferrin splice variant can be that of Genbank AccessionAAA59479. In another specific embodiment, the neutrophil lactoferrinsplice variant can comprise the following amino acid sequenceEDCIALKGEADA (SEQ ID NO: 8), which includes the novel region ofsplice-variance.

In another embodiment, the transferrin portion of the transferrin fusionprotein of the invention includes a melanotransferrin variant.

Modified Tf fusions may be made with any Tf protein, fragment, domain,or engineered domain. For instance, fusion proteins may be producedusing the full-length Tf sequence, with or without the native Tf signalsequence. Tf fusion proteins may also be made using a single Tf domain,such as an individual N or C domain or a modified form of Tf comprising2N or 2C domains (see U.S. Provisional Application 60/406,977, filedAug. 30, 2002, which is herein incorporated by reference in itsentirety). In some embodiments, fusions of a therapeutic protein to asingle C domain may be produced, wherein the C domain is altered toreduce, inhibit or prevent glycosylation. In other embodiments, the useof a single N domain is advantageous as the Tf glycosylation sitesreside in the C domain and the N domain, on its own. A preferredembodiment is the Tf fusion protein having a single N domain which isexpressed at a high level.

As used herein, a C terminal domain or lobe modified to function as anN-like domain is modified to exhibit glycosylation patterns or ironbinding properties substantially like that of a native or wild-type Ndomain or lobe. In a preferred embodiment, the C domain or lobe ismodified so that it is not glycosylated and does not bind iron bysubstitution of the relevant C domain regions or amino acids to thosepresent in the corresponding regions or sites of a native or wild-type Ndomain.

As used herein, a Tf moiety comprising “two N domains or lobes” includesa Tf molecule that is modified to replace the native C domain or lobewith a native or wild-type N domain or lobe or a modified N domain orlobe or contains a C domain that has been modified to functionsubstantially like a wild-type or modified N domain. Analysis of the twodomains by overlay of the two domains (Swiss PDB Viewer 3.7b2, IterativeMagic Fit) and by direct amino acid alignment (ClustalW multiplealignment) reveals that the two domains have diverged over time. Aminoacid alignment shows 42% identity and 59% similarity between the twodomains. However, approximately 80% of the N domain matches the C domainfor structural equivalence. The C domain also has several extradisulfide bonds compared to the N domain.

Alignment of molecular models for the N and C domain reveals thefollowing structural equivalents:

N domain  4-24 36-72  94-136 138-139 149-164 168-173 178-198 219-255259-260 263-268 271-275 279-280 283-288 309-327  (1-330) 75-88 200-214290-304 C domain 340-361 365-415 425-437 470-471 475-490 492-497 507-542555-591 593-594 597-602 605-609 614-615 620-640 645-663 (340-679)439-468The disulfide bonds for the two domains align as follows:

N C C339-C596 C9-C48 C345-C377 C19-C39 C355-C368 C402-C674 C418-C637C118-C194 C450-C523 C137-C331 C474-C665 C158-C174 C484-C498 C161-C179C171-C177 C495-C506 C227-C241 C563-C577 C615-C620 Bold aligned disulfidebonds Italics bridging peptide

In one embodiment, the transferrin portion of the transferrin fusionprotein includes at least two N terminal lobes of transferrin. Infurther embodiments, the transferrin portion of the transferrin fusionprotein includes at least two N terminal lobes of transferrin derivedfrom human serum transferrin.

In another embodiment, the transferrin portion of the transferrin fusionprotein includes, comprises, or consists of at least two C terminallobes of transferrin. In further embodiments, the transferrin portion ofthe transferrin fusion protein includes at least two C terminal lobes oftransferrin derived from human serum transferrin.

In a further embodiment, the C terminal lobe mutant further includes amutation of at least one of Asn413 and Asn611 of SEQ ID NO: 3 which doesnot allow glycosylation.

In some embodiments, the Tf or Tf portion will be of sufficient lengthto increase the in vivo circulatory half-life, serum stability, in vitrosolution stability or bioavailability of the therapeutic proteincompared to the in vivo circulatory half-life, serum stability, in vitrosolution stability or bioavailability of the therapeutic protein in anunfused state. Such an increase in stability, serum half-life orbioavailability may be about a 30%, 50%, 70%, 80%, 90% or more increaseover the unfused therapeutic protein. In some cases, the modifiedtransferrin fusion proteins exhibit a serum half-life of about 10-20 ormore days, about 12-18 days or about 14-17 days.

When the C domain of Tf is part of the fusion protein, the two N-linkedglycosylation sites, amino acid residues corresponding to N413 and N611of SEQ ID NO:3 may be mutated for expression in a yeast system toprevent glycosylation or hypermannosylation and extend the serumhalf-life of the fusion protein and/or therapeutic protein (to produceasialo-, or in some instances, monosialo-Tf or disialo-Tf). In additionto Tf amino acids corresponding to N413 and N611, mutations may be tothe adjacent residues within the N-X-S/T glycosylation site to preventor substantially reduce glycosylation. See U.S. Pat. No. 5,986,067 ofFunk et al. It has also been reported that the N domain of Tf expressedin Pichia pastoris becomes O-linked glycosylated with a single hexose atS32 which also may be mutated or modified to prevent such glycosylation.

Accordingly, in one embodiment of the invention, the transferrin fusionprotein includes a modified transferrin molecule wherein the transferrinexhibits reduced glycosylation, including but not limited toasialo-monosialo- and disialo-forms of Tf. In another embodiment, thetransferrin portion of the transferrin fusion protein includes arecombinant transferrin mutant that is mutated to prevent glycosylation.In another embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant transferrin mutant that is fullyglycosylated. In a further embodiment, the transferrin portion of thetransferrin fusion protein includes a recombinant human serumtransferrin mutant that is mutated to prevent glycosylation, wherein atleast one of Asn413 and Asn611 of SEQ ID NO:3 are mutated to an aminoacid which does not allow glycosylation. In another embodiment, thetransferrin portion of the transferrin fusion protein includes arecombinant human serum transferrin mutant that is mutated to prevent orsubstantially reduce glycosylation, wherein mutations may be to theadjacent residues within the N-X-S/T glycosylation site. Moreover,glycosylation may be reduced or prevented by mutating the serine orthreonine residue. Further, changing the X to proline is known toinhibit glycosylation.

In other embodiments of the invention, the iron binding is retained andthe iron binding ability of Tf may be used to deliver a therapeuticprotein or peptide(s) to the inside of a cell, across an epithelial orendothelial cell membrane and/or across the BBB. These embodiments thatbind iron and/or the Tf receptor will often be engineered to reduce orprevent glycosylation to extend the serum half-life of the therapeuticprotein. The N domain alone will not bind to TfR when loaded with iron,and the iron bound C domain will bind TfR but not with the same affinityas the whole molecule.

In another embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant transferrin mutant having a mutationwherein the mutant does not retain the ability to bind metal ions. In analternate embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant transferrin mutant having a mutationwherein the mutant has a weaker binding avidity for metal ions thanwild-type serum transferrin. In an alternate embodiment, the transferrinportion of the transferrin fusion protein includes a recombinanttransferrin mutant having a mutation wherein the mutant has a strongerbinding avidity for metal ions than wild-type serum transferrin.

In another embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant transferrin mutant having a mutationwherein the mutant does not retain the ability to bind to thetransferrin receptor. In an alternate embodiment, the transferrinportion of the transferrin fusion protein includes a recombinanttransferrin mutant having a mutation wherein the mutant has a weakerbinding avidity for the transferrin receptor than wild-type serumtransferrin. In an alternate embodiment, the transferrin portion of thetransferrin fusion protein includes a recombinant transferrin mutanthaving a mutation wherein the mutant has a stronger binding avidity forthe transferrin receptor than wild-type serum transferrin.

In another embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant transferrin mutant having a mutationwherein the mutant does not retain the ability to bind to carbonateions. In an alternate embodiment, the transferrin portion of thetransferrin fusion protein includes a recombinant transferrin mutanthaving a mutation wherein the mutant has a weaker binding avidity forcarbonate ions than wild-type serum transferrin. In an alternateembodiment, the transferrin portion of the transferrin fusion proteinincludes a recombinant transferrin mutant having a mutation wherein themutant has a stronger binding avidity for carbonate ions than wild-typeserum transferrin.

In another embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant human serum transferrin mutant having amutation in at least one amino acid residue selected from the groupconsisting of Asp63, Gly65, Tyr95, Tyr188, His249, Asp392, Tyr426,Tyr514, Tyr517 and His585 of SEQ ID NO:3, wherein the mutant retains theability to bind metal ions. In an alternate embodiment, a recombinanthuman serum transferrin mutant having a mutation in at least one aminoacid residue selected from the group consisting of Asp63, Gly65, Tyr95,Tyr188, His249, Asp392, Tyr426, Tyr514, Tyr517 and His585 of SEQ IDNO:3, wherein the mutant has a reduced ability to bind metal ions. Inanother embodiment, a recombinant human serum transferrin mutant havinga mutation in at least one amino acid residue selected from the groupconsisting of Asp63, Gly65, Tyr95, Tyr188, His249, Asp392, Tyr426,Tyr517 and His585 of SEQ ID NO:3, wherein the mutant does not retain theability to bind metal ions.

In another embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant human serum transferrin mutant having amutation at Lys206 or His207 of SEQ ID NO:3, wherein the mutant has astronger binding avidity for metal ions than wild-type human serumtransferrin (see U.S. Pat. No. 5,986,067, which is herein incorporatedby reference in its entirety). In an alternate embodiment, thetransferrin portion of the transferrin fusion protein includes arecombinant human serum transferrin mutant having a mutation at Lys206or His207 of SEQ ID NO:3, wherein the mutant has a weaker bindingavidity for metal ions than wild-type human serum transferrin. In afurther embodiment, the transferrin portion of the transferrin fusionprotein includes a recombinant human serum transferrin mutant having amutation at Lys206 or His207 of SEQ ID NO:3, wherein the mutant does notbind metal ions.

Any available technique may be used to make the fusion proteins of theinvention, including but not limited to molecular techniques commonlyavailable, for instance, those disclosed in Sambrook et al. MolecularCloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor LaboratoryPress, 1989. When carrying out nucleotide substitutions using techniquesfor accomplishing site-specific mutagenesis that are well known in theart, the encoded amino acid changes are preferably of a minor nature,that is, conservative amino acid substitutions, although other,non-conservative, substitutions are contemplated as well, particularlywhen producing a modified transferrin portion of a Tf fusion protein,e.g., a modified Tf fusion protein exhibiting reduced glycosylation,reduced iron binding and the like. Specifically contemplated are aminoacid substitutions, small deletions or insertions, typically of one toabout 30 amino acids; insertions between transferrin domains; smallamino- or carboxyl-terminal extensions, such as an amino-terminalmethionine residue, or small linker peptides of less than 50, 40, 30, 20or 10 residues between transferrin domains or linking a transferrinprotein and a therapeutic protein or peptide; or a small extension thatfacilitates purification, such as a poly-histidine tract, an antigenicepitope or a binding domain.

Examples of conservative amino acid substitutions are substitutions madewithin the same group such as within the group of basic amino acids(such as arginine, lysine, histidine), acidic amino acids (such asglutamic acid and aspartic acid), polar amino acids (such as glutamineand asparagine), hydrophobic amino acids (such as leucine, isoleucine,valine), aromatic amino acids (such as phenylalanine, tryptophan,tyrosine) and small amino acids (such as glycine, alanine, serine,threonine, methionine).

Non-conservative substitutions encompass substitutions of amino acids inone group by amino acids in another group. For example, anon-conservative substitution would include the substitution of a polaramino acid for a hydrophobic amino acid. For a general description ofnucleotide substitution, see e.g. Ford et al. (1991), Prot. Exp. Pur. 2:95-107. Non-conservative substitutions, deletions and insertions areparticularly useful to produce TF fusion proteins of the invention thatexhibit no or reduced binding of iron, no or reduced binding of thefusion protein to the Tf receptor.

In the polypeptide and proteins of the invention, the following systemis followed for designating amino acids in accordance with the followingconventional list:

TABLE OF AMINO ACIDS ONE- LETTER THREE-LETTER AMINO ACID SYMBOL SYMBOLAlanine A Ala Arginine R Arg Asparagine N Asn Aspartic Acid D AspCysteine C Cys Glutamine Q Gln Glutamic Acid E Glu Glycine G GlyHistidine H His Isoleucine I Ile Leucine L Leu Lysine K Lys Methionine MMet Phenylalanine F Phe Proline P Pro Serine S Ser Threonine T ThrTryptophan W Trp Tyrosine Y Tyr Valine V Val

Iron binding and/or receptor binding may be reduced or disrupted bymutation, including deletion, substitution or insertion into, amino acidresidues corresponding to one or more of Tf N domain residues Asp63,Tyr95, Tyr188, His249 and/or C domain residues Asp 392, Tyr 426, Tyr 514and/or His 585 of SEQ ID NO: 3. Iron binding may also be affected bymutation to amino acids Lys206, His207 or Arg632 of SEQ ID NO: 3.Carbonate binding may be reduced or disrupted by mutation, includingdeletion, substitution or insertion into, amino acid residuescorresponding to one or more of Tf N domain residues Thr120, Arg124,Ala126, Gly 127 and/or C domain residues Thr 452, Arg 456, Ala 458and/or Gly 459 of SEQ ID NO: 3. A reduction or disruption of carbonatebinding may adversely affect iron and/or receptor binding.

Binding to the Tf receptor may be reduced or disrupted by mutation,including deletion, substitution or insertion into, amino acid residuescorresponding to one or more of Tf N domain residues described above foriron binding.

As discussed above, glycosylation may be reduced or prevented bymutation, including deletion, substitution or insertion into, amino acidresidues corresponding to one or more of Tf C domain residues around theN-X-S/T sites corresponding to C domain residues N413 and/or N611 (SeeU.S. Pat. No. 5,986,067). For instance, the N413 and/or N611 may bemutated to Glu residues.

In instances where the Tf fusion proteins of the invention are notmodified to prevent glycosylation, iron binding, carbonate bindingand/or receptor binding, glycosylation, iron and/or carbonate ions maybe stripped from or cleaved off of the fusion protein. For instance,available deglycosylases may be used to cleave glycosylation residuesfrom the fusion protein, in particular the sugar residues attached tothe Tf portion, yeast deficient in glycosylation enzymes may be used toprevent glycosylation and/or recombinant cells may be grown in thepresence of an agent that prevents glycosylation, e.g., tunicamycin.

The carbohydrates on the fusion protein may also be reduced orcompletely removed enzymatically by treating the fusion protein withdeglycosylases. Deglycosylases are well known in the art. Examples ofdeglycosylases include but are not limited to galactosidase, PNGase A,PNGase F, glucosidase, mannosidase, fucosidase, and Endo Hdeglycosylase.

Nevertheless, in certain circumstances, it may be preferable for oraldelivery that the Tf portion of the fusion protein be fully glycosylated

Additional mutations may be made with Tf to alter the three dimensionalstructure of Tf, such as modifications to the hinge region to preventthe conformational change needed for iron biding and Tf receptorrecognition. For instance, mutations may be made in or around N domainamino acid residues 94-96, 245-247 and/or 316-318 as well as C domainamino acid residues 425-427, 581-582 and/or 652-658. In addition,mutations may be made in or around the flanking regions of these sitesto alter Tf structure and function.

In one aspect of the invention, the transferrin fusion protein canfunction as a carrier protein to extend the half life or bioavailabilityof the therapeutic protein as well as, in some instances, delivering thetherapeutic protein inside a cell and/or across the blood brain barrier.In an alternate embodiment, the transferrin fusion protein includes amodified transferrin molecule wherein the transferrin does not retainthe ability to cross the blood brain barrier.

In another embodiment, the transferrin fusion protein includes amodified transferrin molecule wherein the transferrin molecule retainsthe ability to bind to the transferrin receptor and transport thetherapeutic peptide inside cells. In an alternate embodiment, thetransferrin fusion protein includes a modified transferrin moleculewherein the transferrin molecule does not retain the ability to bind tothe transferrin receptor and transport the therapeutic peptide insidecells.

In further embodiments, the transferrin fusion protein includes amodified transferrin molecule wherein the transferrin molecule retainsthe ability to bind to the transferrin receptor and transport thetherapeutic peptide inside cells and retains the ability to cross theblood brain barrier. In an alternate embodiment, the transferrin fusionprotein includes a modified transferrin molecule wherein the transferrinmolecule retains the ability to cross the blood brain barrier, but doesnot retain the ability to bind to the transferrin receptor and transportthe therapeutic peptide inside cells.

Modified Transferrin Fusion Proteins

The fusion proteins of the invention may contain one or more copies ofthe therapeutic protein or polypeptide attached to the N-terminus and/orthe C-terminus of the Tf protein. In some embodiments, the therapeuticprotein or polypeptide is attached to both the N- and C-terminus of theTf protein and the fusion protein may contain one or more equivalents ofthe therapeutic protein or polypeptide on either or both ends of Tf. Inother embodiments, the therapeutic protein or polypeptide is insertedinto known domains of the Tf protein, for instance, into one or more ofthe loops of Tf (see Ali et al. (1999) J. Biol. Chem.274(34):24066-24073). In other embodiments, the therapeutic protein ortherapeutic peptide is inserted between the N and C domains of Tf.

Generally, the transferrin fusion protein of the invention may have onemodified transferrin-derived region and one therapeutic protein-derivedregion. Multiple regions of each protein, however, may be used to make atransferrin fusion protein of the invention. Similarly, more than onetherapeutic protein may be used to make a transferrin fusion protein ofthe invention, thereby producing a multi-functional modified Tf fusionprotein.

The present invention provides transferrin fusion protein containing atherapeutic protein or polypeptide or portion thereof fused to atransferrin molecule or portion thereof. In one embodiment, thetransferrin fusion protein of the invention contains a therapeuticprotein or polypeptide fused to the N terminus of a transferrinmolecule. In an alternate embodiment, the transferrin fusion protein ofthe invention contains a therapeutic protein fused to the C terminus ofa transferrin molecule. The present invention also provides transferrinfusion protein containing a therapeutic protein or polypeptide orprotion thereof fused to a modified transferrin morlecule or portionthererof.

In other embodiments, the transferrin fusion protein of the inventionscontains a therapeutic protein fused to both the N-terminus and theC-terminus of modified transferrin. In another embodiment, thetherapeutic proteins fused at the N- and C-termini are the sametherapeutic proteins. In an alternate embodiment, the therapeuticproteins fused at the N- and C-termini are different therapeuticproteins. In another alternate embodiment, the therapeutic proteinsfused to the N- and C-termini are different therapeutic proteins whichmay be used to treat or prevent the same disease, disorder, orcondition. In another embodiment, the therapeutic proteins fused at theN- and C-termini are different therapeutic proteins which may be used totreat or prevent diseases or disorders which are known in the art tocommonly occur in patients simultaneously.

In addition to modified transferrin fusion protein of the inventions inwhich the modified transferrin portion is fused to the N terminal and/orC-terminal of the therapeutic protein portion, transferrin fusionprotein of the inventions of the invention may also be produced byinserting the therapeutic protein or peptide of interest (e.g., atherapeutic protein or peptide as disclosed herein, or, for instance, asingle chain antibody that binds a therapeutic protein or a fragment orvariant thereof) into an internal region of the modified transferrin.Internal regions of modified transferrin include, but are not limitedto, the iron binding sites, the hinge regions, the bicarbonate bindingsites, or the receptor binding domain.

Within the protein sequence of the modified transferrin molecule anumber of loops or turns exist, which are stabilized by disulfide bonds.These loops are useful for the insertion, or internal fusion, oftherapeutically active peptides, particularly those requiring asecondary structure to be functional, or therapeutic proteins togenerate a modified transferrin molecule with specific biologicalactivity.

When therapeutic proteins or peptides are inserted into or replace atleast one loop of a Tf molecule, insertions may be made within any ofthe surface exposed loop regions, in addition to other areas of Tf. Forinstance, insertions may be made within the loops comprising Tf aminoacids 32-33, 74-75, 256-257, 279-280 and 288-289 (Ali et al., supra)(See FIG. 3). As previously described, insertions may also be madewithin other regions of Tf such as the sites for iron and bicarbonatebinding, hinge regions, and the receptor binding domain as described inmore detail below. The loops in the Tf protein sequence that areamenable to modification/replacement for the insertion of proteins orpeptides may also be used for the development of a screenable library ofrandom peptide inserts. Any procedures may be used to produce nucleicacid inserts for the generation of peptide libraries, includingavailable phage and bacterial display systems, prior to cloning into aTf domain and/or fusion to the ends of Tf. In other embodiments, thelibrary is made directly in or on the ends of a Tf peptide as describedbelow.

The N-terminus of Tf is free and points away from the body of themolecule. Fusions of proteins or peptides on the N-terminus maytherefore be a preferred embodiment. Such fusions may include a linkerregion, such as but not limited to a poly-glycine stretch, to separatethe therapeutic protein or peptide from Tf. Attention to the junctionbetween the leader sequence, the choice of leader sequence, and thestructure of the mRNA by codon manipulation/optimization (no major stemloops to inhibit ribosome progress) will increase secretion and can bereadily accomplished using standard recombinant protein techniques.

The C-terminus of Tf appears to be more buried and secured by adisulfide bond 6 amino acids from the C-terminus. In human Tf, theC-terminal amino acid is a proline which, depending on the way that itis orientated, will either point a fusion away or into the body of themolecule. A linker or spacer moiety at the C-terminus may be used insome embodiments of the invention. There is also a proline near theN-terminus. In one aspect of the invention, the proline at the N- and/orthe C-termini may be changed out. In another aspect of the invention,the C-terminal disulfide bond may be eliminated to untether theC-terminus.

In yet other embodiments, small molecule therapeutics may be complexedwith iron and loaded on a modified Tf protein fusion for delivery to theinside of cells and across the BBB. The addition of a targeting peptideor, for example, a single chain antibody (SCA) can be used to target thepayload to a particular cell type, e.g., a cancer cell.

Nucleic Acids

Nucleic acid molecules are also provided by the present invention. Theseencode a modified Tf fusion protein comprising a transferrin protein ora portion of a transferrin protein covalently linked or joined to atherapeutic protein. As discussed in more detail below, any therapeuticprotein may be used. The fusion protein may further comprise a linkerregion, for instance a linker less than about 50, 40, 30, 20, or 10amino acid residues. The linker can be covalently linked to and betweenthe transferrin protein or portion thereof and the therapeutic protein.Nucleic acid molecules of the invention may be purified or not.

Host cells and vectors for replicating the nucleic acid molecules andfor expressing the encoded fusion proteins are also provided. Anyvectors or host cells may be used, whether prokaryotic or eukaryotic,but eukaryotic expression systems, in particular yeast expressionsystems, may be preferred. Many vectors and host cells are known in theart for such purposes. It is well within the skill of the art to selectan appropriate set for the desired application.

DNA sequences encoding transferrin, portions of transferrin andtherapeutic proteins of interest may be cloned from a variety of genomicor cDNA libraries known in the art. The techniques for isolating suchDNA sequences using probe-based methods are conventional techniques andare well known to those skilled in the art. Probes for isolating suchDNA sequences may be based on published DNA or protein sequences (see,for example, Baldwin, G. S. (1993) Comparison of Transferrin Sequencesfrom Different Species. Comp. Biochem. Physiol. 106B/1:203-218 and allreferences cited therein, which are hereby incorporated by reference intheir entirety). Alternatively, the polymerase chain reaction (PCR)method disclosed by Mullis et al. (U.S. Pat. No. 4,683,195) and Mullis(U.S. Pat. No. 4,683,202), incorporated herein by reference may be used.The choice of library and selection of probes for the isolation of suchDNA sequences is within the level of ordinary skill in the art.

As known in the art, “similarity” between two polynucleotides orpolypeptides is determined by comparing the nucleotide or amino acidsequence and its conserved nucleotide or amino acid substitutes of onepolynucleotide or polypeptide to the sequence of a second polynucleotideor polypeptide. Also known in the art is “identity” which means thedegree of sequence relatedness between two polypeptide or twopolynucleotide sequences as determined by the identity of the matchbetween two strings of such sequences. Both identity and similarity canbe readily calculated (Computational Molecular Biology, Lesk, A. M.,ed., Oxford University Press, New York, 1988; Biocomputing: Informaticsand Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993;Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin,H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis inMolecular Biology, von Heinje, G., Academic Press, 1987; and SequenceAnalysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press,New York, 1991).

While there exist a number of methods to measure identity and similaritybetween two polynucleotide or polypeptide sequences, the terms“identity” and “similarity” are well known to skilled artisans (SequenceAnalysis in Molecular Biology, von Heinje, G., Academic Press, 1987;Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., MStockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J.Applied Math., 48: 1073 (1988). Methods commonly employed to determineidentity or similarity between two sequences include, but are notlimited to those, disclosed in Guide to Huge Computers, Martin J.Bishop, ed., Academic Press, San Diego, 1994, and Carillo, H., andLipman, D., SIAM J. Applied Math. 48:1073 (1988).

Preferred methods to determine identity are designed to give the largestmatch between the two sequences tested. Methods to determine identityand similarity are codified in computer programs. Preferred computerprogram methods to determine identity and similarity between twosequences include, but are not limited to, GCG program package(Devereux, et al., Nucl. Acid Res. 12(1):387 (1984)), BLASTP, BLASTN,FASTA (Atschul, et al., J. Mol. Biol. 215:403 (1990)). The degree ofsimilarity or identity referred to above is determined as the degree ofidentity between the two sequences, often indicating a derivation of thefirst sequence from the second. The degree of identity between twonucleic acid sequences may be determined by means of computer programsknown in the art such as GAP provided in the GCG program package(Needleman and Wunsch J. Mol. Biol. 48:443-453 (1970)). For purposes ofdetermining the degree of identity between two nucleic acid sequencesfor the present invention, GAP is used with the following settings: GAPcreation penalty of 5.0 and GAP extension penalty of 0.3.

Codon Optimization

The degeneracy of the genetic code permits variations of the nucleotidesequence of a transferrin protein and/or therapeutic protein ofinterest, while still producing a polypeptide having the identical aminoacid sequence as the polypeptide encoded by the native DNA sequence. Theprocedure, known as “codon optimization” (described in U.S. Pat. No.5,547,871 which is incorporated herein by reference in its entirety)provides one with a means of designing such an altered DNA sequence. Thedesign of codon optimized genes should take into account a variety offactors, including the frequency of codon usage in an organism, nearestneighbor frequencies, RNA stability, the potential for secondarystructure formation, the route of synthesis and the intended future DNAmanipulations of that gene. In particular, available methods may be usedto alter the codons encoding a given fusion protein with those mostreadily recognized by yeast when yeast expression systems are used.

The degeneracy of the genetic code permits the same amino acid sequenceto be encoded and translated in many different ways. For example,leucine, serine and arginine are each encoded by six different codons,while valine, proline, threonine, alanine and glycine are each encodedby four different codons. However, the frequency of use of suchsynonymous codons varies from genome to genome among eukaryotes andprokaryotes. For example, synonymous codon-choice patterns among mammalsare very similar, while evolutionarily distant organisms such as yeast(such as S. cerevisiae), bacteria (such as E. coli) and insects (such asD. melanogaster) reveal a clearly different pattern of genomic codon usefrequencies (Grantham, R., et al., Nucl. Acid Res., 8, 49-62 (1980);Grantham, R., et al., Nucl. Acid Res., 9, 43-74 (1981); Maroyama, T., etal., Nucl. Acid Res., 14, 151-197 (1986); Aota, S., et al., Nucl. AcidRes., 16, 315-402 (1988); Wada, K., et al., Nucl. Acid Res., 19 Supp.,1981-1985 (1991); Kurland, C. G., FEBS Lett., 285, 165-169 (1991)).These differences in codon-choice patterns appear to contribute to theoverall expression levels of individual genes by modulating peptideelongation rates. (Kurland, C. G., FEBS Lett., 285, 165-169 (1991);Pedersen, S., EMBO J., 3, 2895-2898 (1984); Sorensen, M. A., J. Mol.Biol., 207, 365-377 (1989); Randall, L. L., et al., Eur. J. Biochem.,107, 375-379 (1980); Curran, J. F., and Yarus, M., J. Mol. Biol., 209,65-77 (1989); Varenne, S., et al., J. Mol. Biol., 180, 549-576 (1984),Varenne, S., et al., J. Mol, Biol., 180, 549-576 (1984); Garel, J.-P.,J. Theor. Biol., 43, 211-225 (1974); Ikemura, T., J. Mol. Biol., 146,1-21 (1981); Ikemura, T., J. Mol. Biol., 151, 389-409 (1981)).

The preferred codon usage frequencies for a synthetic gene shouldreflect the codon usages of nuclear genes derived from the exact (or asclosely related as possible) genome of the cell/organism that isintended to be used for recombinant protein expression, particularlythat of yeast species. As discussed above, in one preferred embodimentthe human Tf sequence is codon optimized, before or after modificationas herein described for yeast expression as may be the therapeuticprotein nucleotide sequence(s).

Vectors

Expression units for use in the present invention will generallycomprise the following elements, operably linked in a 5′ to 3′orientation: a transcriptional promoter, a secretory signal sequence, aDNA sequence encoding a modified Tf fusion protein comprisingtransferrin protein or a portion of a transferrin protein joined to aDNA sequence encoding a therapeutic protein or peptide of interest and atranscriptional terminator. As discussed above, any arrangement of thetherapeutic protein or peptide fused to or within the Tf portion may beused in the vectors of the invention. The selection of suitablepromoters, signal sequences and terminators will be determined by theselected host cell and will be evident to one skilled in the art and arediscussed more specifically below.

Suitable yeast vectors for use in the present invention are described inU.S. Pat. No. 6,291,212 and include YRp7 (Struhl et al., Proc. Natl.Acad. Sci. USA 76: 1035-1039, 1978), YEp13 (Broach et al., Gene 8:121-133, 1979), pJDB249 and pJDB219 (Beggs, Nature 275:104-108, 1978),pPPC0005, pSeCHSA, pScNHSA, pC4 and derivatives thereof. Useful yeastplasmid vectors also include pRS403-406, pRS413-416 and the Pichiavectors available from Stratagene Cloning Systems, La Jolla, Calif.92037, USA. Plasmids pRS403, pRS404, pRS405 and pRS406 are YeastIntegrating plasmids (YIps) and incorporate the yeast selectable markersHIS3, TRP1, LEU2 and URA3. PlasmidspRS413˜41.6 are Yeast Centromereplasmids (YCps).

Such vectors will generally include a selectable marker, which may beone of any number of genes that exhibit a dominant phenotype for which aphenotypic assay exists to enable transformants to be selected.Preferred selectable markers are those that complement host cellauxotrophy, provide antibiotic resistance or enable a cell to utilizespecific carbon sources, and include LEU2 (Broach et al. ibid.), URA3(Botstein et al., Gene 8: 17, 1979), HIS3 (Struhl et al., ibid.) or POT1(Kawasaki and Bell, EP 171,142). Other suitable selectable markersinclude the CAT gene, which confers chloramphenicol resistance on yeastcells. Preferred promoters for use in yeast include promoters from yeastglycolytic genes (Hitzeman et al., J Biol. Chem. 225: 12073-12080, 1980;Alber and Kawasaki, J. Mol. Appl. Genet. 1: 419-434, 1982; Kawasaki,U.S. Pat. No. 4,599,311) or alcohol dehydrogenase genes (Young et al.,in Genetic Engineering of Microorganisms for Chemicals, Hollaender etal., (eds.), p. 355, Plenum, N.Y., 1982; Ammerer, Meth. Enzymol. 101:192-201, 1983). In this regard, particularly preferred promoters are theTPI1 promoter (Kawasaki, U.S. Pat. No. 4,599,311) and the ADH2-4^(C)(see U.S. Pat. No. 6,291,212 promoter (Russell et al., Nature 304:652-654, 1983). The expression units may also include a transcriptionalterminator. A preferred transcriptional terminator is the TPI1terminator (Alber and Kawasaki, ibid.). Other preferred vectors andpreferred components such as promoters and terminators of a yeastexpression system are disclosed in European Patents EP 0258067, EP0286424, EP0317254, EP 0387319, EP 0386222, EP 0424117, EP 0431880, andEP 1002095; European Patent Publications EP 0828759, EP 0764209, EP0749478, and EP 0889949; PCT Publication WO 00/44772 and WO 94/04687;and U.S. Pat. Nos. 5,739,007; 5,637,504; 5,302,697; 5,260,202;5,667,986; 5,728,553; 5,783,423; 5,965,386; 6150,133; 6,379,924; and5,714,377; which are herein incorporated by reference in their entirety.

In addition to yeast, modified fusion proteins of the present inventioncan be expressed in filamentous fungi, for example, strains of the fungiAspergillus. Examples of useful promoters include those derived fromAspergillus nidulans glycolytic genes, such as the adh3 promoter(McKnight et al., EMBO J. 4: 2093-2099, 1985) and the tpiA promoter. Anexample of a suitable terminator is the adh3 terminator (McKnight etal., ibid.). The expression units utilizing such components may becloned into vectors that are capable of insertion into the chromosomalDNA of Aspergillus, for example.

Mammalian expression vectors for use in carrying out the presentinvention will include a promoter capable of directing the transcriptionof the modified Tf fusion protein. Preferred promoters include viralpromoters and cellular promoters. Preferred viral promoters include themajor late promoter from adenovirus 2 (Kaufman and Sharp, Mol. Cell.Biol. 2: 1304-13199, 1982) and the SV40 promoter (Subramani et al., Mol.Cell. Biol. 1: 854-864, 1981). Preferred cellular promoters include themouse metallothionein 1 promoter (Palmiter et al., Science 222: 809-814,1983) and a mouse Vκ (see U.S. Pat. No. 6,291,212) promoter (Grant etal., Nuc. Acids Res. 15: 5496, 1987). A particularly preferred promoteris a mouse V_(H) (see U.S. Pat. No. 6,291,212) promoter (Loh et al.,ibid.). Such expression vectors may also contain a set of RNA splicesites located downstream from the promoter and upstream from the DNAsequence encoding the transferrin fusion protein. Preferred RNA splicesites may be obtained from adenovirus and/or immunoglobulin genes.

Also contained in the expression vectors is a polyadenylation signallocated downstream of the coding sequence of interest. Polyadenylationsignals include the early or late polyadenylation signals from SV40(Kaufman and Sharp, ibid.), the polyadenylation signal from theadenovirus 5 E1B region and the human growth hormone gene terminator(DeNoto et al., Nucl. Acid Res. 9: 3719-3730, 1981). A particularlypreferred polyadenylation signal is the V_(H) (see U.S. Pat. No.6,291,212) gene terminator (Loh et al., ibid.). The expression vectorsmay include a noncoding viral leader sequence, such as the adenovirus 2tripartite leader, located between the promoter and the RNA splicesites. Preferred vectors may also include enhancer sequences, such asthe SV40 enhancer and the mouse (see U.S. Pat. No. 6,291,212) enhancer(Gillies, Cell 33: 717-728, 1983). Expression vectors may also includesequences encoding the adenovirus VA RNAs.

Transformation

Techniques for transforming fungi are well known in the literature, andhave been described, for instance, by Beggs (ibid.), Hinnen et al.(Proc. Natl. Acad. Sci. USA 75: 1929-1933, 1978), Yelton et al., (Proc.Natl. Acad. Sci. USA 81: 1740-1747, 1984), and Russell (Nature 301:167-169, 1983). Other techniques for introducing cloned DNA sequencesinto fungal cells, such as electroporation (Becker and Guarente, Methodsin Enzymol. 194: 182-187, 1991) may be used. The genotype of the hostcell will generally contain a genetic defect that is complemented by theselectable marker present on the expression vector. Choice of aparticular host and selectable marker is well within the level ofordinary skill in the art.

Cloned DNA sequences comprising modified Tf fusion proteins of theinvention may be introduced into cultured mammalian cells by, forexample, calcium phosphate-mediated transfection (Wigler et al., Cell14: 725, 1978; Corsaro and Pearson, Somatic Cell Genetics 7: 603, 1981;Graham and Van der Eb, Virology 52: 456, 1973.) Other techniques forintroducing cloned DNA sequences into mammalian cells, such aselectroporation (Neumann et al., EMBO J. 1: 841-845, 1982), orlipofection may also be used. In order to identify cells that haveintegrated the cloned DNA, a selectable marker is generally introducedinto the cells along with the gene or cDNA of interest. Preferredselectable markers for use in cultured mammalian cells include genesthat confer resistance to drugs, such as neomycin, hygromycin, andmethotrexate. The selectable marker may be an amplifiable selectablemarker. A preferred amplifiable selectable marker is the DHFR gene. Aparticularly preferred amplifiable marker is the DHFR^(r) (see U.S. Pat.No. 6,291,212) cDNA (Simonsen and Levinson, Proc. Natl. Acad. Sci. USA80: 2495-2499, 1983). Selectable markers are reviewed by Thilly(Mammalian Cell Technology, Butterworth Publishers, Stoneham, Mass.) andthe choice of selectable markers is well within the level of ordinaryskill in the art.

Host Cells

The present invention also includes a cell, preferably a yeast celltransformed to express a modified transferrin fusion protein of theinvention. In addition to the transformed host cells themselves, thepresent invention also includes a culture of those cells, preferably amonoclonal (clonally homogeneous) culture, or a culture derived from amonoclonal culture, in a nutrient medium. If the polypeptide issecreted, the medium will contain the polypeptide, with the cells, orwithout the cells if they have been filtered or centrifuged away.

Host cells for use in practicing the present invention includeeukaryotic cells, and in some cases prokaryotic cells, capable of beingtransformed or transfected with exogenous DNA and grown in culture, suchas cultured mammalian, insect, fungal, plant and bacterial cells.

Fungal cells, including species of yeast (e.g., Saccharomyces spp.,Schizosaccharomyces spp., Pichia spp.) may be used as host cells withinthe present invention. Examples of fungi including yeasts contemplatedto be useful in the practice, of the present invention as hosts forexpressing the transferrin fusion protein of the inventions are Pichia(some species of which were formerly classified as Hansenula),Saccharomyces, Kluyveromyces spergillus, Candida, Torulopsis,Torulaspora, Schizosaccharomyces, Citeromyces, Pachysolen,Zygosaccharomyces, Debaromyces, Trichoderma, Cephalosporium, Humicola,Mucor, Neurospora, Yarrowia, Metschunikowia, Rhodosporidium,Leucosporidiunz, Botryoascus, Sporidiobolus, Endomycopsis, and the like.Examples of Saccharomyces spp. are S. cerevisiae, S. italicus and S.rouxii. Examples of Kluyveromyces spp. are K. fragilis, K. lactis and K.marxianus. A suitable Torulaspora species is T. delbrueckii. Examples ofPichia spp. are P. angusta (formerly H. polymorpha), P. anomala(formerly H. anomala) and P. pastoris.

Particularly useful host cells to produce the Tf fusion proteins of theinvention are the methylotrophic Pichia pastoris (Steinlein et al.(1995) Protein Express. Purif. 6:619-624). Pichia pastoris has beendeveloped to be an outstanding host for the production of foreignproteins since its alcohol oxidase promoter was isolated and cloned; itstransformation was first reported in 1985. P. pastoris can utilizemethanol as a carbon source in the absence of glucose. The P. pastorisexpression system can use the methanol-induced alcohol oxidase (AOX1)promoter, which controls the gene that codes for the expression ofalcohol oxidase, the enzyme which catalyzes the first step in themetabolism of methanol. This promoter has been characterized andincorporated into a series of P. pastoris expression vectors. Since theproteins produced in P. pastoris are typically folded correctly andsecreted into the medium, the fermentation of genetically engineered P.pastoris provides an excellent alternative to E. coli expressionsystems. A number of proteins have been produced using this system,including tetanus toxin fragment, Bordatella pertussis pertactin, humanserum albumin and lysozyme.

Strains of the yeast Saccharomyces cerevisiae are another preferredhost. In a preferred embodiment, a yeast cell, or more specifically, aSaccharomyces cerevisiae host cell that contains a genetic deficiency ina gene required for asparagine-linked glycosylation of glycoproteins isused. S. cerevisiae host cells having such defects may be prepared usingstandard techniques of mutation and selection, although many availableyeast strains have been modified to prevent or reduce glycosylation orhypermannosylation. Ballou et al. (J. Biol. Chem. 255: 5986-5991, 1980)have described the isolation of mannoprotein biosynthesis mutants thatare defective in genes which affect asparagine-linked glycosylation.Gentzsch and Tanner (Glycobiology 7:481-486, 1997) have described afamily of at least six genes (PMT1-6) encoding enzymes responsible forthe first step in O-glycosylation of proteins in yeast. Mutantsdefective in one or more of these genes show reduced O-linkedglycosylation and/or altered specificity of O-glycosylation.

To optimize production of the heterologous proteins, it is alsopreferred that the host strain carries a mutation, such as the S.cerevisiae pep4 mutation (Jones, Genetics 85: 23-33, 1977), whichresults in reduced proteolytic activity. Host strains containingmutations in other protease encoding regions are particularly useful toproduce large quantities of the Tf fusion proteins of the invention.

Host cells containing DNA constructs of the present invention are grownin an appropriate growth medium. As used herein, the term “appropriategrowth medium” means a medium containing nutrients required for thegrowth of cells. Nutrients required for cell growth may include a carbonsource, a nitrogen source, essential amino acids, vitamins, minerals andgrowth factors. The growth medium will generally select for cellscontaining the DNA construct by, for example, drug selection ordeficiency in an essential nutrient which is complemented by theselectable marker on the DNA construct or co-transfected with the DNAconstruct. Yeast cells, for example, are preferably grown in achemically defined medium, comprising a carbon source, e.g. sucrose, anon-amino acid nitrogen source, inorganic salts, vitamins and essentialamino acid supplements. The pH of the medium is preferably maintained ata pH greater than 2 and less than 8, preferably at pH 5.5-6.5. Methodsfor maintaining a stable pH include buffering and constant pH control.Preferred buffering agents include succinic acid and Bis-Tris (SigmaChemical Co., St. Louis, Mo.). Yeast cells having a defect in a generequired for asparagine-linked glycosylation are preferably grown in amedium containing an osmotic stabilizer. A preferred osmotic stabilizeris sorbitol supplemented into the medium at a concentration between 0.1M and 1.5 M., preferably at 0.5 M or 1.0 M.

Cultured mammalian cells are generally grown in commercially availableserum-containing or serum-free media. Selection of a medium appropriatefor the particular cell line used is within the level of ordinary skillin the art. Transfected mammalian cells are allowed to grow for a periodof time, typically 1-2 days, to begin expressing the DNA sequence(s) ofinterest. Drug selection is then applied to select for growth of cellsthat are expressing the selectable marker in a stable fashion. For cellsthat have been transfected with an amplifiable selectable marker thedrug concentration may be increased in a stepwise manner to select forincreased copy number of the cloned sequences, thereby increasingexpression levels.

Baculovirus/insect cell expression systems may also be used to producethe modified Tf fusion proteins of the invention. The BacPAK™Baculovirus Expression System (BD Biosciences (Clontech)) expressesrecombinant proteins at high levels in insect host cells. The targetgene is inserted into a transfer vector, which is cotransfected intoinsect host cells with the linearized BacPAK6 viral DNA. The BacPAK6 DNAis missing an essential portion of the baculovirus genome. When the DNArecombines with the vector, the essential element is restored and thetarget gene is transferred to the baculovirus genome. Followingrecombination, a few viral plaques are picked and purified, and therecombinant phenotype is verified. The newly isolated recombinant viruscan then be amplified and used to infect insect cell cultures to producelarge amounts of the desired protein.

Tf fusion proteins of the present invention may also be produced usingtransgenic plants and animals. For example, sheep and goats can make thetherapeutic protein in their milk. Or tobacco plants can include theprotein in their leaves. Both transgenic plant and animal production ofproteins comprises adding a new gene coding the fusion protein into thegenome of the organism. Not only can the transgenic organism produce anew protein, but it can also pass this ability onto its offspring.

Secretory Signal Sequences

The terms “secretory signal sequence” or “signal sequence” or “secretionleader sequence” are used interchangeably and are described, for examplein U.S. Pat. No. 6,291,212 and U.S. Pat. No. 5,547,871, both of whichare herein incorporated by reference in their entirety. Secretory signalsequences or signal sequences or secretion leader sequences encodesecretory peptides. A secretory peptide is an amino acid sequence thatacts to direct the secretion of a mature polypeptide or protein from acell. Secretory peptides are generally characterized by a core ofhydrophobic amino acids and are typically (but not exclusively) found atthe amino termini of newly synthesized proteins. Very often thesecretory peptide is cleaved from the mature protein during secretion.Secretory peptides may contain processing sites that allow cleavage ofthe signal peptide from the mature protein as it passes through thesecretory pathway. Processing sites may be encoded within the signalpeptide or may be added to the signal peptide by, for example, in vitromutagenesis.

Secretory peptides may be used to direct the secretion of modified Tffusion proteins of the invention. One such secretory peptide that may beused in combination with other secretory peptides is the alpha matingfactor leader sequence. Secretory signal sequences or signal sequencesor secretion leader sequences are required for a complex series ofpost-translational processing steps which result in secretion of aprotein. If an intact signal sequence is present, the protein beingexpressed enters the lumen of the rough endoplasmic reticulum and isthen transported through the Golgi apparatus to secretory vesicles andis finally transported out of the cell. Generally, the signal sequenceimmediately follows the initiation codon and encodes a signal peptide atthe amino-terminal end of the protein to be secreted. In most cases, thesignal sequence is cleaved off by a specific protease, called a signalpeptidase. Preferred signal sequences improve the processing and exportefficiency of recombinant protein expression using viral, mammalian oryeast expression vectors. In some cases, the native Tf signal sequencemay be used to express and secrete fusion proteins of the invention.

Linkers

The Tf moiety and therapeutic protein moiety(s) of the modifiedtransferrin fusion proteins of the invention can be fused directly orusing a linker peptide of various lengths to provide greater physicalseparation and allow more spatial mobility between the fused proteinsand thus maximize the accessibility of the therapeutic protein portion,for instance, for binding to its cognate receptor. The linker peptidemay consist of amino acids that are flexible or more rigid. For example,a linker such as but not limited to a poly-glycine stretch. The linkercan be less than about 50, 40, 30, 20, or 10 amino acid residues. Thelinker can be covalently linked to and between the transferrin proteinor portion thereof and the therapeutic protein.

Detection of Tf Fusion Proteins

Assays for detection of biologically active modifiedtransferrin-therapeutic protein fusions may include Western transfer,protein blot or colony filter as well as activity based assays thatdetect the fused therapeutic protein. A Western transfer filter may beprepared using the method described by Towbin et al. (Proc. Natl. Acad.Sci. USA 76: 4350-4354, 1979). Briefly, samples are electrophoresed in asodium dodecylsulfate polyacrylamide gel. The proteins in the gel areelectrophoretically transferred to nitrocellulose paper. Protein blotfilters may be prepared by filtering supernatant samples or concentratesthrough nitrocellulose filters using, for example, a Minifold(Schleicher & Schuell, Keene, N. H.). Colony filters may be prepared bygrowing colonies on a nitrocellulose filter that has been laid across anappropriate growth medium. In this method, a solid medium is preferred.The cells are allowed to grow on the filters for at least 12 hours. Thecells are removed from the filters by washing with an appropriate bufferthat does not remove the proteins bound to the filters. A preferredbuffer comprises 25 mM Tris-base, 19 mM glycine, pH 8.3, 20% methanol.

Fusion proteins of the invention may also be detected by assaying forthe activity of the therapeutic protein moiety. Such assays are readilyavailable, including but not limited to, those assays described inTable 1. Specifically, transferrin fusion proteins of the invention maybe assayed for functional activity (e.g., biological activity ortherapeutic activity) using the assay referenced in the “ExemplaryActivity Assay” column of Table 1. Additionally, one of skill in the artmay routinely assay fragments of a therapeutic protein corresponding toa therapeutic protein portion of a fusion protein of the invention, foractivity using assays referenced in its corresponding row of Table 1.Further, one of skill in the art may routinely assay fragments of amodified transferrin protein for activity using assays known in the art.

For example, in one embodiment where one is assaying for the ability ofa transferrin fusion protein of the invention to bind or compete with atherapeutic protein for binding to an anti-therapeutic polypeptideantibody and/or anti-transferrin antibody, various immunoassays known inthe art can be used, including but not limited to, competitive andnon-competitive assay systems using techniques such asradioimmunoassays, ELISA (enzyme linked immunosorbent assay), sandwichimmunoassays, immunoradiometric assays, gel diffusion precipitationreactions, immunodiffusion assays, in situ immunoassays (using colloidalgold, enzyme or radioisotope labels, for example), western blots,precipitation reactions, agglutination assays (e.g., gel agglutinationassays), complement fixation assays, immunofluorescence assays, proteinA assays, and immunoelectrophoresis assays, etc. In one embodiment,antibody binding is detected by detecting a label on the primaryantibody. In another embodiment, the primary antibody is detected bydetecting binding of a secondary antibody or reagent to the primaryantibody. In a further embodiment, the secondary antibody is labeled.Many means are known in the art for detecting binding in an immunoassayand are within the scope of the present invention.

In a further embodiment, where a binding partner (e.g., a receptor or aligand) of a therapeutic protein is identified, binding to that bindingpartner by a transferrin fusion protein containing that therapeuticprotein as the therapeutic protein portion of the fusion can be assayed,e.g., by means well-known in the art, such as, for example, reducing andnon-reducing gel chromatography, protein affinity chromatography, andaffinity blotting. Other methods will be known to the skilled artisanand are within the scope of the invention.

Isolation/Purification of Modified Transferrin Fusion Proteins

Secreted, biologically active, modified transferrin fusion proteins maybe isolated from the medium of host cells grown under conditions thatallow the secretion of the biologically active fusion proteins. The cellmaterial is removed from the culture medium, and the biologically activefusion proteins are isolated using isolation techniques known in theart. Suitable isolation techniques include precipitation andfractionation by a variety of chromatographic methods, including gelfiltration, ion exchange chromatography and affinity chromatography.

A particularly preferred purification method is affinity chromatographyon an iron binding or metal chelating column or an immunoaffinitychromatography using an antibody directed against the transferrin ortherapeutic protein or peptide portion of the polypeptide fusion. Theantibody is preferably immobilized or attached to a solid support orsubstrate. A particularly preferred substrate is CNBr-activatedSepharose (Pharmacia LKB Technologies, Inc., Piscataway, N.J.). By thismethod, the medium is combined with the antibody/substrate underconditions that will allow binding to occur. The complex may be washedto remove unbound material, and the transferrin fusion protein isreleased or eluted through the use of conditions unfavorable to complexformation. Particularly useful methods of elution include changes in pH,wherein the immobilized antibody has a high affinity for the ligand at afirst pH and a reduced affinity at a second (higher or lower) pH;changes in concentration of certain chaotropic agents; or through theuse of detergents.

Labeled Modified Transferrin Fusion Proteins

Transferrin fusion proteins of the present invention may also be labeledwith a radioisotope or other imaging agent and used for in vivodiagnostic purposes. Preferred radioisotope imaging agents includeiodine-125 and technetium-99, with technetium-99 being particularlypreferred. Methods for producing protein-isotope conjugates are wellknown in the art, and are described by, for example, Eckelman et al.(U.S. Pat. No. 4,652,440), Parker et al. (WO 87/05030) and Wilber et al.(EP 203,764). Alternatively, the transferrin fusion proteins may bebound to spin label enhancers and used for magnetic resonance (MR)imaging. Suitable spin label enhancers include stable, stericallyhindered, free radical compounds such as nitroxides. Methods forlabeling ligands for MR imaging are disclosed by, for example, Coffmanet al. (U.S. Pat. No. 4,656,026). For administration, the labeledtransferrin fusion proteins are combined with a pharmaceuticallyacceptable carrier or diluent, such as sterile saline or sterile water.Administration is preferably by bolus injection, preferablyintravenously.

Production of Fusion Proteins

The present invention further provides methods for producing a modifiedfusion protein of the invention using nucleic acid molecules hereindescribed. In general terms, the production of a recombinant form of aprotein typically involves the following steps.

A nucleic acid molecule is first obtained that encodes a transferrinfusion protein of the invention. The nucleic acid molecule is thenpreferably placed in operable linkage with suitable control sequences,as described above, to form an expression unit containing the proteinopen reading frame. The expression unit is used to transform a suitablehost and the transformed host is cultured under conditions that allowthe production of the recombinant protein. Optionally the recombinantprotein is isolated from the medium or from the cells; recovery andpurification of the protein may not be necessary in some instances wheresome impurities may be tolerated.

Each of the foregoing steps can be accomplished in a variety of ways.For example, the construction of expression vectors that are operable ina variety of hosts is accomplished using appropriate replicons andcontrol sequences, as set forth above. The control sequences, expressionvectors, and transformation methods are dependent on the type of hostcell used to express the gene and were discussed in detail earlier andare otherwise known to persons skilled in the art. Suitable restrictionsites can, if not normally available, be added to the ends of the codingsequence so as to provide an excisable gene to insert into thesevectors. A skilled artisan can readily adapt any host/expression systemknown in the art for use with the nucleic acid molecules of theinvention to produce a desired recombinant protein.

As discussed above, any expression system may be used, including yeast,bacterial, animal, plant, eukaryotic and prokaryotic systems. In someembodiments, yeast, mammalian cell culture and transgenic animal orplant production systems are preferred. In other embodiments, yeastsystems that have been modified to reduce native yeast glycosylation,hyper-glycosylation or proteolytic activity may be used.

Therapeutic Molecules

Any therapeutic molecule may be used as the fusion partner to Tfaccording to the methods and compositions of the present invention. Asused herein, a therapeutic molecule is typically a protein or peptidecapable of exerting a beneficial biological effect in vitro or in vivoand includes proteins or peptides that exert a beneficial effect inrelation to normal homeostasis, physiology or a disease state.Therapeutic molecules do not include, fusion partners commonly used asmarkers or protein purification aids, such as bacterial galactosidases(see for example, U.S. Pat. No. 5,986,067 and Aldred et al. (1984)Biochem. Biophys. Res. Commun. 122: 960-965). For instance, a beneficialeffect as related to a disease state includes any effect that isadvantageous to the treated subject, including disease prevention,disease stabilization, the lessening or alleviation of disease symptomsor a modulation, alleviation or cure of the underlying defect to producean effect beneficial to the treated subject.

A modified transferrin fusion protein of the invention includes at leasta fragment or variant of a therapeutic protein and at least a fragmentor variant of modified serum transferrin, which are associated with oneanother, preferably by genetic fusion.

In one embodiment, the transferrin fusion protein includes a modifiedtransferrin molecule linked to a neuropharmaceutical agent. In anotherembodiment, the modified transferrin fusion protein includes transferrinat the carboxyl terminus linked to a neuropharmaceutical agent at theamino terminus. In an alternate embodiment, the modified transferrinfusion protein includes transferrin at the amino terminus linked to aneuropharmaceutical agent at the carboxy terminus. In specificembodiments, the neuropharmaceutical agent is either nerve growth factoror ciliary neurotrophic factor.

In further embodiments, a modified transferrin fusion protein of theinvention may contain at least a fragment or variant of a therapeuticprotein, and/or at least a fragment or variant of an antibody. In afurther embodiment, the transferrin fusion proteins can contain peptidefragments or peptide variants of proteins or antibodies wherein thevariant or fragment retains at least one biological or therapeuticactivity. The transferrin fusion proteins can contain therapeuticproteins that can be peptide fragments or peptide variants at leastabout 4, at least 5, at least 6, at least 7, at least 8, at least 9, atleast 10, at least 11, at least 12, at least 13, at least 14, at least15, at least 20, at least 25, at least 30, at least 35, or at leastabout 40, at least about 50, at least about 55, at least about 60 or atleast about 70 or more amino acids in length fused to the N and/or Ctermini, inserted within, or inserted into a loop of a modifiedtransferrin.

In another embodiment, the modified transferrin fusion molecules containa therapeutic protein portion that can be fragments of a therapeuticprotein that include the full length protein as well as polypeptideshaving one or more residues deleted from the amino terminus of the aminoacid sequence.

In another embodiment, the modified transferrin fusion molecules containa therapeutic protein portion that can be fragments of a therapeuticprotein that include the full length protein as well as polypeptideshaving one or more residues deleted from the carboxy terminus of theamino acid sequence.

In another embodiment, the modified transferrin fusion molecules containa therapeutic protein portion that can have one or more amino acidsdeleted from both the amino and the carboxy termini.

In another embodiment, the modified transferrin fusion molecules containa therapeutic protein portion that is at least about 80%, 85%, 90%, 95%,96%, 97%, 98% or 99% identical to a reference therapeutic protein setforth herein, or fragments thereof. In further embodiments, thetransferrin fusion molecules contain a therapeutic protein portion thatis at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical toreference polypeptides having the amino acid sequence of N- andC-terminal deletions as described above.

In another embodiment, the modified transferrin fusion molecules containthe therapeutic protein portion that is at least about 80%, 85%, 90%,95%, 96%, 97%, 98%, 99% or 100%, identical to, for example, the nativeor wild-type amino acid sequence of a therapeutic protein. Fragments, ofthese polypeptides are also provided.

The therapeutic proteins corresponding to a therapeutic protein portionof a modified transferrin fusion protein of the invention, such as cellsurface and secretory proteins, can be modified by the attachment of oneor more oligosaccharide groups. The modification referred to asglycosylation can significantly affect the physical properties ofproteins and can be important in protein stability, secretion, andlocalization. Glycosylation occurs at specific locations along thepolypeptide backbone. There are usually two major types ofglycosylation: glycosylation characterized by O-linked oligosaccharides,which are attached to serine or threonine residues; and glycosylationcharacterized by N-linked oligosaccharides, which are attached toasparagine residues in an Asn-X-Ser/Thr sequence, where X can be anamino acid except proline. Variables such as protein structure and celltype influence the number and nature of the carbohydrate units withinthe chains at different glycosylation sites. Glycosylation isomers arealso common at the same site within a given cell type. For example,several types of human interferon are glycosylated.

Therapeutic proteins corresponding to a therapeutic protein portion of atransferrin fusion protein of the invention, as well as analogs andvariants thereof, may be modified so that glycosylation at one or moresites is altered as a result of manipulation(s) of their nucleic acidsequence by the host cell in which they are expressed, or due to otherconditions of their expression. For example, glycosylation isomers maybe produced by abolishing or introducing glycosylation sites, e.g., bysubstitution or deletion of amino acid residues, such as substitution ofglutamine for asparagine, or unglycosylated recombinant proteins may beproduced by expressing the proteins in host cells that will notglycosylate them, e.g. in glycosylation-deficient yeast. Theseapproaches are known in the art.

Therapeutic proteins and their nucleic acid sequences are well known inthe art and available in public databases such as Chemical AbstractsServices Databases (e.g., the CAS Registry), GenBank, and GenSeq. TheAccession Numbers and sequences referred to below are hereinincorporated by reference in their entirety.

In other embodiments, the transferrin fusion proteins of the inventionare capable of a therapeutic activity and/or biologic activity,corresponding to the therapeutic activity and/or biologic activity ofthe therapeutic protein listed in the corresponding row of Table 1 andelsewhere in this application. (See, e.g., the “Biological Activity” and“Therapeutic Protein X” columns of Table 1.) In further embodiments, thetherapeutically active protein portions of the transferrin fusionproteins of the invention are fragments or variants of the referencesequences cited herein.

The present invention is further directed to modified Tf fusion proteinscomprising fragments of the therapeutic proteins herein described. Evenif deletion of one or more amino acids from the N-terminus of a proteinresults in modification or loss of one or more biological functions ofthe therapeutic protein portion, other therapeutic activities and/orfunctional activities (e.g., biological activities, ability tomultimerize, ability to bind a ligand) may still be retained. Forexample, the ability of polypeptides with N-terminal deletions to induceand/or bind to antibodies which recognize the complete or mature formsof the polypeptides generally will be retained with less than themajority of the residues of the complete polypeptide removed from theN-terminus. Whether a particular polypeptide lacking N-terminal residuesof a complete polypeptide retains such immunologic activities can beassayed by routine methods described herein and otherwise known in theart. It is not unlikely that a mutant with a large number of deletedN-terminal amino acid residues may retain some biological or immunogenicactivities. In fact, peptides composed of as few as six amino acidresidues may often evoke an immune response.

Also as mentioned above, even if deletion of one or more amino acidsfrom the N-terminus or C-terminus of a therapeutic protein results inmodification or loss of one or more biological functions of the protein,other functional activities (e.g., biological activities, ability tomultimerize, ability to bind a ligand) and/or therapeutic activities maystill be retained. For example the ability of polypeptides withC-terminal deletions to induce and/or bind to antibodies which recognizethe complete or mature forms of the polypeptide generally will beretained when less than the majority of the residues of the complete ormature polypeptide are removed from the C-terminus. Whether a particularpolypeptide lacking the N-terminal and/or, C-terminal residues of areference polypeptide retains therapeutic activity can readily bedetermined by routine methods described herein and/or otherwise known inthe art.

Peptide fragments of the therapeutic proteins can be fragmentscomprising, or alternatively, consisting of, an amino acid sequence thatdisplays a therapeutic activity and/or functional activity (e.g.biological activity) of the polypeptide sequence of the therapeuticprotein of which the amino acid sequence is a fragment.

The peptide fragments of the therapeutic protein may comprise only theN- and C-termini of the protein, i.e., the central portion of thetherapeutic protein has been deleted. Alternatively, the peptidefragments may comprise non-adjacent and/or adjacent portions of thecentral part of the therapeutic protein.

Other polypeptide fragments are biologically active fragments.Biologically active fragments are those exhibiting activity similar, butnot necessarily identical, to an activity of a therapeutic protein usedin the present invention. The biological activity of the fragments mayinclude an improved desired activity, or a decreased undesirableactivity.

Generally, variants of proteins are overall very similar, and, in manyregions, identical to the amino acid sequence of the therapeutic proteincorresponding to a therapeutic protein portion of a transferrin fusionprotein of the invention. Nucleic acids encoding these variants are alsoencompassed by the invention.

Further therapeutic polypeptides that may be used in the invention arepolypeptides encoded by polynucleotides which hybridize to thecomplement of a nucleic acid molecule encoding an amino-acid sequence ofa therapeutic protein under stringent hybridization conditions which areknown to those of skill in the art. (see, for example, Ausubel, F. M. etal., eds., 1989 Current protocol in Molecular Biology, Green PublishingAssociates, Inc., and John Wiley & Sons Inc., New. York).Polynucleotides encoding these polypeptides are also encompassed by theinvention.

By a polypeptide-having an amino acid sequence at least, for example,95% “identical” to a query amino acid sequence of the present invention,it is intended that the amino acid sequence of the subject polypeptideis identical to the query sequence except that the subject polypeptidesequence may include up to five amino acid alterations per each 100amino acids of the query amino acid sequence. In other words, to obtaina polypeptide having an amino acid sequence at least 95% identical to aquery amino acid sequence, up to 5% of the amino acid residues in thesubject sequence may be inserted, deleted, or substituted with anotheramino acid. These alterations of the reference sequence may occur at theamino- or carboxy-terminal positions of the reference amino acidsequence or anywhere between those terminal positions, interspersedeither individually among residues in the reference sequence, or in oneor more contiguous groups within the reference sequence.

As a practical matter, whether any particular polypeptide is at leastabout 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to, forinstance, the amino acid sequence of a transferrin fusion protein of theinvention or a fragment thereof (such, as the therapeutic proteinportion of the transferrin fusion protein or the transferrin portion ofthe transferrin fusion protein), can be determined conventionally usingknown computer programs. A preferred method for determining the bestoverall match between a query sequence (a sequence of the presentinvention) and a subject sequence, also referred to as a global sequencealignment, can be determined using the FASTDB computer program based onthe algorithm of Brufiag et al. (Comp. App. Biosci 245 (1990)).

The polynucleotide variants of the invention may contain alterations inthe coding regions, non-coding regions, or both. Polynucleotide variantscontaining alterations which produce silent substitutions, additions, ordeletions, but do not alter the properties or activities of the encodedpolypeptide may be used to produce modified Tf fusion proteins.Nucleotide variants produced by silent substitutions due to thedegeneracy of the genetic code can be utilized. Moreover, polypeptidevariants in which less than about 50, less than 40, less than 30, lessthan 20, less than 10, or 5-50, 5-25, 5-10, 1-5, or 1-2 amino acids aresubstituted, deleted, or added in any combination can also be utilized.Polynucleotide variants can be produced for a variety of reasons, e.g.,to optimize codon expression for a particular host (change codons in thehuman mRNA to those preferred by a host, such as, yeast or E. coli asdescribed above).

In other embodiments, the therapeutic protein moiety has conservativesubstitutions compared to the wild-type sequence. By “conservativesubstitutions” is intended swaps within groups such as replacement ofthe aliphatic or hydrophobic amino acids Ala, Val, Leu and Ile;replacement of the hydroxyl residues Ser and Thr; replacement of theacidic residues Asp and Glu; replacement of the amide residues Asn andGln, replacement of the basic residues Lys, Arg, and His; replacement ofthe aromatic residues Phe, Tyr, and Trp, and replacement of thesmall-sized amino acids Ala, Ser, Thr, Met, and Gly. Guidance concerninghow to make phenotypically silent amino acid substitutions is provided,for example, in Bowie et al., “Deciphering the Message in ProteinSequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310(1990). In specific embodiments, the polypeptides of the inventioncomprise, or alternatively, consist of, fragments or variants of theamino acid sequence of a therapeutic protein described herein and/orserum transferrin, and/modified transferrin protein of the invention,wherein the fragments or variants have 1-5, 5-10, 5-25, 5-50, 10-50 or50-150 amino acid residue additions, substitutions, and/or deletionswhen compared to the reference amino acid sequence. In furtherembodiments, the amino acid substitutions are conservative. Nucleicacids encoding these polypeptides are also encompassed by the invention.

The modified fusion proteins of the present invention can be composed ofamino-acids joined to each other by peptide bonds or modified peptidebonds and may contain amino acids other than the 20 gene-encoded aminoacids. The polypeptides may be modified by either natural processes,such as post-translational processing, or by chemical modificationtechniques which are well known in the art. Such modifications are welldescribed in basic texts and in more detailed monographs, as well as ina voluminous research literature.

Modifications can occur anywhere in a polypeptide, including the peptidebackbone, the amino acid side-chains and the amino or carboxy termini.It will be appreciated that the same type of modification may be presentin the same or varying degrees at several sites in a given polypeptide.Also, a given polypeptide may contain many types of modifications.Polypeptides may be branched, for example, as a result ofubiquitination, and they may be cyclic, with or without branching.Cyclic, branched, and branched cyclic polypeptides may result fromposttranslation natural processes or may be made by synthetic methods.Modifications include acetylation, acylation, ADP-ribosylation,amidation, covalent attachment of flavin, covalent attachment of a hememoiety, covalent attachment of a nucleotide or nucleotide derivative,covalent attachment of a lipid or lipid derivative, covalent attachmentof phosphotidylinositol, cross-linking, cyclization, disulfide bondformation, demethylation, formation of covalent cross-links, formationof cysteine, glycosylation, GPI anchor formation, hydroxylation,iodination, methylation, myristylation, oxidation, pegylation,proteolytic processing, phosphorylation, prenylation, racemization,sulfation, transfer-RNA mediated addition of amino acids to proteinssuch as arginylation, and ubiquitination. (See, for instance,PROTEINS—STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton,W. H. Freeman and Company, New York (1993); POST-TRANSLATIONAL COVALENTMODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York,pgs. 1-12 (1983); Seifter et al. (1990) Meth. Enzymol. 182:626-646;Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62.

Therapeutic molecules that may be fused to or inserted into Tf include,but are not limited to, hormones, matrix proteins, immunosuppressants,bronchodilators, cardiovascular agents, enzymes, CNS agents,neurotransmitters, receptor proteins or peptides, growth hormones,growth factors, antiviral peptides, fusogenic inhibitor peptides,cytokines, lymphokines, monokines, interleukins, colony stimulatingfactors, differentiation factors, angiogenic factors, receptor ligands,cancer-associated proteins, antineoplastics, viral peptides, antibioticpeptides, blood proteins, antagonist proteins, transcription factors,anti-angiogenic factors, antagonist proteins or peptides, receptorantagonists, antibodies, single chain antibodies and cell adhesionmolecules. Different therapeutic molecules may be combined into a singlefusion protein to produce a bi or multi-functional therapeutic molecule.Different molecules may also be used in combination to produce a fusionprotein with a therapeutic entity and a targeting entity.

Cytokines are soluble proteins released by cells of the immune system,which act nonenzymatically through specific receptors to regulate immuneresponses. Cytokines resemble hormones in that they act at lowconcentrations bound with high affinity to a specific receptor. The term“cytokine” is used herein to describe naturally occurring or recombinantproteins, analogs thereof, and fragments thereof which elicit a specificbiological response in a cell which has a receptor for that cytokine.Cytokines preferably include interleukins such as interleukin-2 (IL-2)(GenBank Acc. No. S77834), IL-3 (GenBank Acc. No. M14743), IL-4 (GenBankAcc. No. M23442), IL-5 (GenBank Acc. No. J03478), IL-6 (GenBank Acc. No.M14584), IL-7 (GenBank Acc. No. NM_(—)000880), IL-10 (GenBank Acc. No.NM_(—)000572), IL-12 (GenBank Acc. No. AF180562 and GenBank Acc. No.AF180563), IL-13 (GenBank Acc. No. U10307), IL-14 (GenBank Acc. No.XM_(—)170924), IL-15 (GenBank Acc. No. X91233), IL-16 (GenBank Acc. No.NM_(—)004513), IL-17 (GenBank Acc. No. NM_(—)002190) and IL-18 (GenBankAcc. No. NM_(—)001562), hematopoietic factors such asgranulocyte-macrophage colony stimulating factor (GM-CSF) (GenBank Acc.No. X03021), granulocyte colony stimulating factor (G-CSF) (GenBank Acc.No. X03656), platelet activating factor (GenBank Acc. No. NM_(—)000437)and erythropoietin (GenBank Acc. No. X02158), tumor necrosis factors(TNF) such as TNFα (GenBank Acc. No. X02910), lymphokines such aslymphotoxin-α (GenBank Acc. No. X02911), lymphotoxin-β (GenBank Acc. No.L11016), leukoregulin, macrophage migration inhibitory factor (GenBankAcc. No. M25639), and neuroleukin (GenBank Acc. No. K03515), regulatorsof metabolic processes such as leptin (GenBank Acc. No. U43415),interferons such as interferon α (IFNα) (GenBank Acc. No. M54886), IFNβ(GenBank Acc. No. V00534), IFNγ (GenBank Acc. No. J00219), IFNα (GenBankAcc. No. NM_(—)002177), thrombospondin 1 (THBS1) (GenBank Acc. No.NM_(—)003246), THBS2 (GenBank Acc. No. L12350), THBS3 (GenBank Acc. No.L38969), THBS4 (GenBank Acc. No. NM_(—)003248), and chemokines.Preferably, the modified transferrin-cytokine fusion protein of thepresent invention displays cytokine biological activity.

The term “hormone” is used herein to describe any one of a number ofbiologically active substances that are produced by certain cells ortissues and that cause specific biological changes or activities tooccur in another cell or tissue located elsewhere in the body. Hormonespreferably include GLP-1 of glucagon preproprotein (GenBank Acc. No.NM_(—)002045), proinsulin (GenBank Acc. No. V00565), insulin (GenBankAcc. No. NM_(—)000207), growth hormone 1 (GenBank Acc. No. V00520),growth hormone 2 (GenBank Acc. No. F006060), growth hormone releasefactor (GenBank Acc. No. NM_(—)021081), insulin-like growth factor I(GenBank Acc. No. M27544), insulin-like growth factor II (GenBank Acc.No. NM_(—)000612), insulin-like growth factor binding protein 1(IGFBP-1) (GenBank Acc. No. M59316), IGFBP-2 (GenBank Acc. No. X16302),IGFBP-3 (GenBank Acc. No. NM_(—)000598), IGFBP-4 (GenBank Acc. No.Y12508), IGFBP-5 (GenBank Acc. No. M65062), IGFBP-6 (GenBank Acc. No.NM_(—)002178), IGFBP-7 (GenBank Acc. No. NM_(—)001553), chorionicgonadotropin 13 chain (GenBank Acc. No. NM_(—)033142), chorionicgonadotropin a chain (GenBank Acc. No. NM_(—)000735), luteinizinghormone β (GenBank Acc. No. X00264), follicle-stimulating hormone β(GenBank Acc. No. NM_(—)000510), thyroid-stimulating hormone β (GenBankAcc. No. NM_(—)000549), prolactin (GenBank Acc. No. NM_(—)000948),pro-opiomelanocortin (GenBank Acc. No. V01510), corticotropin (ACTH),β-lipotropin, α-melanocyte stimulating hormone (α-MSH), γ-lipotropin,β-MSH, β-endorphin, and corticotropin-like intermediate lobe peptide(CLIP).

The term “growth factor” is used herein to describe any protein orpeptide that binds to a receptor to stimulate cell proliferation. Growthfactors preferably include platelet-derived growth factor-α (PDGF-α)(GenBank Acc. No. X03795); PDGF-β (GenBank Acc. No. X02811), steroidhormones, epidermal growth factor (EGF) (GenBank Acc. No. NM_(—)001963),fibroblast growth factors such as fibroblast growth factor 1 (FGF1)(GenBank Acc. No. NM_(—)000800), FGF2 (GenBank Acc. No. NM_(—)002006),FGF3 (GenBank Acc. No. NM_(—)005247), FGF4 (GenBank Acc. No.NM_(—)002007), FGF5 (GenBank Acc. No. M37825), FGF6 (GenBank Acc. No.X57075), FGF7 (GenBank Acc. No. NM_(—)002009), FGF8 (GenBank Acc. No.AH006649), FGF9 (GenBank Acc. No. NM_(—)002010), FGF10 (GenBank Acc. No.AB002097), FGF11 (GenBank Acc. No. NM_(—)004112), FGF12 (GenBank Acc.No. NM_(—)021032), FGF13 (GenBank Acc. No. NM_(—)004114), FGF14 (GenBankAcc. No. NM_(—)004115), FGF16 (GenBank Acc. No. AB009391), FGF17(GenBank Acc. No. NM_(—)003867), FGF18 (GenBank Acc. No. AF075292),FGF19 (GenBank Acc. No. NM_(—)005117), FGF20 (GenBank Acc. No.NM_(—)019851), FGF21 (GenBank Acc. No. NM_(—)019113), FGF22 (GenBankAcc. No. NM_(—)020637), and FGF23 (GenBank Acc. No. NM_(—)020638),angiogenin (GenBank Acc. No. M11567), brain-derived neurotrophic factor(GenBank Acc. No. M61176), ciliary neurotrophic growth factor (GenBankAcc. No. X60542), transforming growth factor-α (TGF-α) (GenBank Acc. No.X70340), TGF-β (GenBank Acc. No. X02812), nerve growth factor-α (NGF-α)(GenBank Acc. No. NM_(—)010915), NGF-β (GenBank Acc. No. X52599), tissueinhibitor of metalloproteinase 1 (TIMP1) (GenBank Acc. No.NM_(—)003254), TIMP2 (GenBank Acc. No. NM_(—)003255), TIMP3 (GenBankAcc. No. U02571), TIMP4 (GenBank Acc. No. U76456) and macrophagestimulating 1 (GenBank Acc. No. L11924).

The term “matrix protein” is used herein to describe proteins orpeptides that are normally found in the extracellular matrix. Theseproteins may be functionally important for strength, filtration, oradhesion. Matrix proteins preferably include collagens such as collagenI (GenBank Acc. No. Z74615), collagen II (GenBank Acc. No. X16711),collagen III (GenBank Acc. No. X14420), collagen IV (GenBank Acc. No.NM_(—)001845), collagen V (GenBank Acc. No. NM_(—)000393), collagen VI(GenBank Acc. No. NM_(—)058175), collagen VII (GenBank Acc. No. L02870),collagen VIII (GenBank Acc. No. NM_(—)001850), collagen IX (GenBank Acc.No. X54412), collagen X (GenBank Acc. No. X60382), collagen XI (GenBankAcc. No. J04177), and collagen XII (GenBank Acc. No. U73778), lamininproteins such as LAMA2 (GenBank Acc. No. NM_(—)000426), LAMA3 (GenBankAcc. No. L34155), LAMA4 (GenBank Acc. No. NM_(—)002290), LAMB1 (GenBankAcc. No. NM_(—)002291), LAMB3 (GenBank Acc. No. L25541), LAMC1 (GenBankAcc. No. NM_(—)002293), nidogen (GenBank Acc. No. NM_(—)002508),α-tectorin (GenBank Acc. No. NM_(—)005422), β-tectorin (GenBank Acc. No.NM_(—)058222), and fibronectin (GenBank Ace. No. X02761).

The term “blood proteins” are traditionally defined as those sourcedfrom plasma, many now commonly produced by recombinant means, andinclude, but are not limited to native serum proteins, derivatives,fragments and mutants or variants thereof, blood clotting factors,derivatives, mutants, variants and fragments (including factors VII,VIII, IX, X), protease inhibitors (antithrombin 3, alpha-1 antitrypsin),urokinase-type plasminogen activator, immunoglobulins, von Willebrandfactor and von Willebrand mutants, fibronectin, fibrinogen, thrombin andhemoglobin.

The term “enzyme” is used herein to describe any protein orproteinaceous substance which catalyzes a specific reaction withoutitself being permanently altered or destroyed. Enzymes preferablyinclude coagulation factors such as F2 (GenBank Acc. No. XM_(—)170688),F7 (GenBank Acc. No. XM_(—)027508), F8 (GenBank Acc. No. XM_(—)013124),F9 (GenBank Acc. No. NM_(—)000133), F10 (GenBank Acc. No. AF503510) andothers, matrix metalloproteinases such as matrix metalloproteinase I(GenBank Acc. No. MMP1) (GenBank Acc. No. NM_(—)002421), MMP2 (GenBankAcc. No. NM_(—)004530), MMP3 (GenBank Acc. No. NM_(—)002422), MMP7(GenBank Acc. No. NM_(—)002423), MMP8 (GenBank Acc. No. NM_(—)002424),MMP9 (GenBank Acc. No. NM_(—)004994), MMP10 (GenBank Acc. No.NM_(—)002425), MMP12 (GenBank Acc. No. NM_(—)002426), MMP13 (GenBankAcc. No. X75308), MMP20 (GenBank Acc. No. NM_(—)004771), adenosinedeaminase (GenBank Acc. No. NM_(—)000022), mitogen activated proteinkinases such as MAPK3 (GenBank Acc. No. XM_(—)055766), MAP2K2 (GenBankAcc. No. NM_(—)030662), MAP2K1 (GenBank Acc. No. NM_(—)002755), MAP2K4(GenBank Acc. No. NM_(—)003010), MAP2K7 (AF013588), and MAPK12(NM_(—)002969), kinases such as JNKK1 (GenBank Acc. No. U17743), JNKK2(GenBank Acc. No. AF014401), JAK1 (M64174), JAK2 (NM_(—)004972), andJAK3 (NM_(—)000215), and phosphatases such as PPM1A (GenBank Acc. No.NM_(—)021003) and PPM1D (GenBank Acc. No. NM_(—)003620).

The term “transcription factors” is used herein to describe any proteinor peptide involved in the transcription of protein-coding genes.Transcription factors may include Sp1, Sp2 (GenBank Acc. No.NM_(—)003110), Sp3 (GenBank Acc. No. AY070137), Sp4 (GenBank Acc. No.NM_(—)003112) NFYB (GenBank Acc. No. NM_(—)006166), Hap2 (GenBank Acc.No. M59079), GATA-1 (GenBank Acc. No. NM_(—)002049), GATA-2 (GenBankAcc. No. NM_(—)002050), GATA-3 (GenBank Acc. No. X55122), GATA-4(GenBank Acc. No. L34357), GATA-5, GATA-6 (GenBank Acc. No.NM_(—)005257), FOG2 (NM_(—)012082), Eryf1 (GenBank Acc. No. X17254),TRPS1 (GenBank Acc. No. NM_(—)014112), NF-E2 (GenBank Acc. No.NM_(—)006163), NF-E3, NF-E4, TFCP2 (GenBank Acc. No. NM_(—)005653),Oct-1 (GenBank Acc. No. X13403), homeobox proteins such as HOXB2(GenBank Acc. No. NM_(—)002145), HOX2H (GenBank Acc. No. X16665),hairless homolog (GenBank Acc. No. NM_(—)005144), mothers againstdecapentaplegic proteins such as MADH1 (GenBank Acc. No. NM_(—)005900),MADH2 (GenBank Acc. No. NM_(—)005901), MADH3 (GenBank Acc. No.NM_(—)005902), MADH4 (GenBank Acc. No. NM_(—)005359), MADH5 (GenBankAcc. No. AF009678), MADH6 (GenBank Acc. No. NM_(—)005585), MADH7(GenBank Acc. No. NM_(—)005904), MADH9 (GenBank Acc. No. NM_(—)005905),and signal transducer and activator of transcription proteins such asSTAT1 (GenBank Acc. No. XM_(—)010893), STAT2 (GenBank Acc. No.NM_(—)005419), STAT3 (GenBank Acc. No. AJ012463), STAT4 (GenBank Acc.No. NM_(—)003151), STAT5 (GenBank Acc. No. L41142), and STAT6 (GenBankAcc. No. NM_(—)003153).

In yet another embodiment of the invention, the therapeutic molecule isa non-human or non-mammalian protein. For example, HIV gp120, HIV Tat,surface proteins of other viruses such as hepatitis, herpes, influenza,adenovirus and RSV, other HIV components, parasitic surface proteinssuch as malarial antigens, and bacterial surface proteins are preferred.These non-human proteins may be used, for example, as antigens, orbecause they have useful activities. For example, the therapeuticmolecule may be streptokinase, staphylokinase, asparaginase, or otherproteins with useful enzymatic activities.

In an alternative embodiment, the therapeutic molecule is aligand-binding protein with biological activity. Such ligand-bindingproteins may, for example, (1) block receptor-ligand interactions at thecell surface; or (2) neutralize the biological activity of a molecule inthe fluid phase of the blood, thereby preventing it from reaching itscellular target. In some embodiments, the modified transferrin fusionproteins include a modified transferrin molecule fused to aligand-binding domain of a receptor selected from the group consistingof, but not limited to, a low density lipoprotein (LDL) receptor, anacetylated LDL receptor, a tumor necrosis factor α receptor, atransforming growth factor β receptor, a cytokine receptor, animmunoglobulin Fc receptor, a hormone receptor, a glucose receptor, aglycolipid receptor, and a glycosaminoglycan receptor. In otherembodiments, ligand-binding proteins include CD2 (M14362), CD3G(NM_(—)000073), CD3D (NM_(—)000732), CD3E (NM_(—)000733), CD3Z (J04132),CD28 (NM_(—)006139), CD4 (GenBank Acc. No. NM_(—)000616), CD1A (GenBankAcc. No. M28825), CD1B (GenBank Acc. No. NM_(—)001764), CD1C (GenBankAcc. No. NM_(—)001765), CD1D (GenBank Acc. No. NM_(—)001766), CD80(GenBank Acc. No. NM_(—)005191), GNB3 (GenBank Acc. No. AF501884),CTLA-4 (GenBank Acc. No. NM_(—)005214), intercellular adhesion moleculessuch as ICAM-1 (NM_(—)000201), ICAM-2 (NM_(—)000873), and ICAM-3(NM_(—)002162), tumor necrosis factor receptors such as TNFRSF1A(GenBank Acc. No. X55313), TNFR1SFB (GenBank Acc. No. NM_(—)001066),TNFRSF9 (GenBank Acc. No. NM_(—)001561), TNFRSF10B (GenBank Acc. No.NM_(—)003842), TNFRSF11B (GenBank Acc. No. NM_(—)002546), and TNFRSF13B(GenBank Acc. No. NM_(—)006573), and interleukin receptors such as IL2RA(GenBank Acc. No. NM_(—)000417), IL2RG (GenBank Acc. No. NM_(—)000206),IL4R (GenBank Acc. No. AF421855), IL7R (GenBank Acc. No. NM_(—)002185),IL9R (GenBank Acc. No. XM_(—)015989), and IL13R (GenBank Acc. No.X95302). Preferably, the Tf-ligand-binding protein fusion of the presentinvention displays the biological activity of the ligand-bindingprotein.

The term “cancer-associated proteins” is used herein to describeproteins or polypeptides whose expression is associated with cancer orthe maintenance of controlled cell growth, such as proteins encoded bytumor suppressor genes or oncogenes. Cancer-associated proteins mayinclude p16 (GenBank Acc. No. AH005371), p53 (GenBank Acc. No.NM_(—)000546), p63 (GenBank Acc. No. NM_(—)003722), p73 (GenBank Acc.No. NM_(—)05427), BRCA1 (GenBank Acc. No. U14680), BRCA2 (GenBank Acc.No. NM_(—)000059), CTBP interacting protein (GenBank Acc. No. U72066),DMBT1 (GenBank Acc. No. NM_(—)004406), HRAS (GenBank Acc. No.NM_(—)005343), NCYM (GenBank Acc. No. NM_(—)006316), FGR (GenBank Acc.No. NM_(—)005248), myb (GenBank Acc. No. AF104863), raf1 (GenBank Acc.No. NM_(—)002880), erbB2 (GenBank Acc. No. NM_(—)004448), VAV (GenBankAcc. No. X16316), c-fos (V GenBank Acc. No. 01512), c-fes (GenBank Acc.No. X52192), c-jun (GenBank Acc. No. NM_(—)002228), MAS1 (GenBank Acc.No. M13150), pim-1 (GenBank Acc. No. M16750), TIF1 (GenBank Acc. No.NM_(—)003852), c-fms (GenBank Acc. No. X03663), EGFR (GenBank Acc. No.NM_(—)005228), erbA (GenBank Acc. No. X04707), c-src tyrosine kinase(GenBank Acc. No. XM_(—)044659), c-abl (GenBank Acc. No. M14752), N-ras(GenBank Acc. No. X02751), K-ras (GenBank Acc. No. M54968), jun-B(GenBank Ace. No. M29039), c-myc (GenBank Acc. No. AH001511), RB1(GenBank Acc. No. M28419), DCC (GenBank Acc. No. X76132), APC (GenBankAcc. No. NM_(—)000038), NF1 (GenBank Acc. No. M89914), NF2 (GenBank Acc.No. Y18000), and bcl-2 (GenBank Acc. No. M13994).

“Fusogenic inhibitor peptides” is used herein to describe peptides thatshow antiviral activity, anti-membrane fusion capability, and/or anability to modulate intracellular processes, for instance, thoseinvolving coiled-coil peptide structures. Antiviral activity includes,but is not limited to, the inhibition of HIV-1, HIV-2, RSV, SIV, EBV,measles virus, influenza virus, or CMV transmission to uninfected cells.Additionally, the antifusogenic capability, antiviral activity orintracellular modulatory activity of the peptides merely requires thepresence of the peptides and specifically does not require thestimulation of a host immune response directed against such peptides.Antifusogenic refers to a peptide's ability to inhibit or reduce thelevel of membrane fusion events between two or more moieties relative tothe level of membrane fusion which occurs between said moieties in theabsence of the peptide. The moieties may be, for example, cell membranesor viral structures, such as viral envelopes or pili. The term“antiviral peptide”, as used herein, refers to the peptide's ability toinhibit viral infection of cells or some viral activity required forproductive viral infection and/or viral pathogenesis, via, for example,cell-cell fusion or free virus infection. Such infection may involvemembrane fusion, as occurs in the case of enveloped viruses, or someother fusion event involving a viral structure and a cellular structure.Fusogenic inhibitor peptides and antiviral peptides often have aminoacid sequences that are derived from greater than one viral protein(e.g., an HIV-1, HIV-2, RSV, and SIV-derived polypeptide).

Examples of fusogenic inhibitor peptides and antiviral peptides can befound in WO 94/2820, WO 96/19495, WO 96/40191, WO 01/64013 and U.S. Pat.Nos. 6,333,395, 6,258,782, 6,228,983, 6,133,418, 6,093,794, 6,068,973,6,060,065, 6,054,265, 6,020,459, 6,017,536, 6,013,263, 5,464,933,5,346,989, 5,603,933, 5,656,480, 5,759,517, 6,245,737; 6,326,004, and6,348,568; all of which are herein incorporated by reference. In apreferred embodiment, antifusogenic peptides are selected from the groupconsisting of HIV T-20 (FWNWLSAWKDLELLEQENKEQQNQSEEILSHILSTY, SEQ ID NO:4), HIV T-1249, RSV T786 (VYPSDEYDASISQVNEEINQALAYIRKADELLENV, SEQ IDNO: 5), RSV T1584 (AVSKVLHLEGEVNKIKSALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQL,SEQ ID NO: 6) and RSV T112 (VFPSDEFDASISQVNEKINQSLAFIRESDELLHNV, SEQ IDNO: 7).

Examples of other types of peptides, include fragments of therapeuticproteins as described herein, in particular, fragments of human proteinsthat retain at least one activity of the parent molecule. Peptides thatmay be used to produce modified Tf fusion proteins of the invention alsoinclude mimetic peptides and peptides that exhibit a biological activityof a therapeutic protein but differ in sequence or three-dimensionalstructure from a full-length therapeutic protein. As a non-limitedexample, peptides include erythropoietin mimetic peptides disclosed byJohnson et al. (2000) Nephrol. Dial. Transplant 15(9): 1274-7, Kuai etal. (2000) J. Pept. Res. 56(2):59-62, Barbone et al. (1999) Nephrol.Dial. Transplant. 14 Supp 2:80-4, Middleton et al. (1999) J. Biol. Chem.274(20):14163-9, Johnson et al. (1998) Biochemistry 37(10:3699-710,Johnson et al. (1997) Chem. Biol. 12:939-50, Wrighton et al. (1997) Nat.Biotechnol. 15(12):1261-5, Livnah et al. (1996) Science 273:464-71, andWrighton et al., (1996) Science 273:458-64.

Therapeutic molecules also include allergenic proteins and digestedfragments thereof. These include pollen allergens from ragweed, rye,June grass, orchard grass, sweet vernal grass, red top grass, timothygrass, yellow dock, wheat, corn, sagebrush, blue grass, Californiaannual grass, pigweed, Bermuda grass, Russian thistle, mountain cedar,oak, box elder, sycamore, maple, elm, etc., dust mites, bee venom, foodallergens, animal dander, and other insect venoms.

Other therapeutic molecules include microbial vaccines which includeviral, bacterial and protozoal vaccines and their various componentssuch as surface antigens. These include vaccines which containglycoproteins, proteins or peptides derived from these proteins. Suchvaccines are prepared from Staphylococcus aureus, Streptococcuspyogenes, Streptococcus pneumoniae, Neisseria meningitidis, Neisseriagonorrhoeae, Salmonella spp., Shigella spp., Escherichia coli,Klebsiella spp., Proteus spp., Vibrio cholerae, Campylobacter pylori,Pseudomonas aeruginosa, Haemophilus influenzae, Bordetella pertussis,Mycobacterium tuberculosis, Legionella pneumophila, Treponema pallidum,chlamydia, tetanus toxoid, diphtheria toxoid, influenza viruses,adenoviruses, paramyxoviruses (mumps, measles), rubella viruses, polioviruses, hepatitis viruses, herpes viruses, rabies virus, HIV-1, HIV-2,RSV and papilloma viruses.

Preferred fusion molecules may contain anti-HIV viral peptides, anti-RSVpeptides, human growth hormone, a and/or J3 interferons, erythropoietin(EPO), EPO like peptides, granulocyte-colony stimulating factor (GCSF),granulocyte-macrophage colony-stimulating factor (GMCSF), insulin,insulin-like growth factor (IGF), thrombopoeitin, peptides correspondingto the CDR of an antibody, Islet Neogenesis Associated Protein (INGAP),calcitonin, angiostatin, endostatin, interleukin-2, growth hormonereleasing factor, human parathyroid hormone, anti-tumor necrosis factor(TNF) peptides, interleukin-1 (IL-1) receptor and/or single chainantibodies.

Fusion proteins of the invention may also be prepared to includepeptides or polypeptides derived from peptide libraries to screen formolecules with new or novel functions. Such peptide libraries mayinclude those commercially or publicly available, e.g., American PeptideCo. Inc., Cell Sciences Inc., Invitrogen Corporation, PhoenixPharmaceuticals Inc., United States Biological, as well as thoseproduced by available technologies, e.g., bacteriophage and bacterialdisplay libraries made using standard procedures.

In yet other embodiments of the invention, Tf fusion proteins may beprepared by using therapeutic protein moieties as known in the art andexemplified by the peptides and proteins currently approved by the Foodand Drug Administration at (www.fda.gov/cber/efoi/approve.htm) as wellas PCT Patent Publication Nos. WO 01/79258, WO 01/77137, WO 01/79442, WO01/79443, WO 01/79444 and WO 01/79480, all of which are hereinincorporated by reference in their entirety.

Table 1 (adapted from PCT International Publication No. WO 01/79444)provides a non-exhaustive list of therapeutic proteins that correspondto a therapeutic protein portion of a modified transferrin fusionprotein of the invention. The “Therapeutic Protein X” column disclosestherapeutic protein molecules followed by parentheses containingscientific and brand names that comprise or alternatively consist ofthat therapeutic protein molecule or a fragment or variant thereof.“Therapeutic protein X” as used herein may refer either to an individualtherapeutic protein molecule (as defined by the amino acid sequenceobtainable from the CAS and Genbank accession numbers), or to the entiregroup of therapeutic proteins associated with a given therapeuticprotein molecule disclosed in this column. The ‘Exemplary Identifier’column provides Chemical Abstracts Services (CAS) Registry Numbers(published by the American Chemical Society) and/or Genbank AccessionNumbers (e.g., Locus ID, NP-XXXXX (Reference Sequence Protein), andXP-XXXXX (Model Protein) identifiers available through the NationalCenter for Biotechnology Information (NCBI) webpage(www.ncbi.nlm.nih.gov) that correspond to entries in the CAS Registry orGenbank database which contain an amino acid sequence of the proteinmolecule or of a fragment or variant of the therapeutic proteinmolecule. In addition GenSeq Accession numbers and/or journalpublication citations are given to identify the exemplary amino acidsequence for some polypeptides.

The summary pages associated with each of these CAS and Genbank andGenSeq Accession Numbers as well as the cited journal publications areavailable (e.g., PubMed ID number (PMID)) and are herein incorporated byreference in their entirety. The PCT/Patent Reference column providesU.S. Patent numbers, or PCT International Publication Numberscorresponding to patents and/or published patent-applications thatdescribe the therapeutic protein molecule all of which are hereinincorporated by reference in their entirety. The Biological Activitycolumn describes biological activities associated with the therapeuticprotein molecule. The Exemplary Activity Assay column providesreferences that describe assays which may be used to test thetherapeutic and/or biological activity of a therapeutic protein or atransferrin fusion protein of the invention comprising a therapeuticprotein X portion. These references are also herein incorporated byreference in their entirety. “The Preferred Indication Y” columndescribes disease, disorders, and/or conditions that may be treated,prevented, diagnosed, or ameliorated by therapeutic protein X or atransferrin fusion protein of the invention comprising a therapeuticprotein X portion.

TABLE 1 Therapeutic Exemplary PCT/Patent Preferred Protein X IdentifierReference Biological Activity Exemplary Activity Assay Indication YBMP-1 GeneSeq WO8800205 BMP1 belongs to the transforming BMP-1 activitycan be determined Induction of Cartilage, Acession P80618 growthfactor-beta (TGFB) superfamily. using the following assays known inTissue and Bone Bone morphogenic proteins induce the art: Nat Genet.2001 Growth, and Diabetes cartilage and bone formation, play January;27(1): 84-8; Eur J Biochem 1996 important role in nephrogesis, and playApr. 1; 237(1): 295-302; J Biol Chem, an important role in thedevelopment of Vol. 274, Issue 16, 10897-10902, many organs, includinglung, heart, Apr. 16, 1999; and Hogan, B. L. M. teeth, gut, skin, andparticularly the (1996) Genes Dev. 10, 1580-1594. kidney. BMP-2 GeneSeqWO8800205 BMP-2 belongs to the transforming BMP-2 activity can bedetermined Induction of Cartilage, Accession P80619 growth factor-beta(TGFB) superfamily. using the following assays known in Tissue and BoneBone morphogenic protein induces bone the art: Nat Genet. 2001 January;Growth, and Diabetes formation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1;237(1): 295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16,1999; and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. BMP-2BGeneSeq U.S. Pat. No. 5,631,142 BMP-2b belongs to the transformingBMP-2b activity can be determined Induction of Cartilage, Accessiongrowth factor-beta (TGFB) superfamily. using the following assays knownin Tissue and Bone W24850 Bone morphogenic protein induces bone the art:Nat Genet. 2001 January; Growth, and Diabetes formation. 27(1): 84-8;Eur J Biochem 1996 Apr. 1; 237(1): 295-302; I Biol Cbcre, Vol. 274,Issue 16, 10897-10902, Apr. 16, 1999; and Hogan, B. L. M. (1996) GenesDev. 10, 1580-1594. BMP-4 GeneSeq WO0020591 BMP-4 belongs to thetransforming BMP-4 activity can be determined Induction of Cartilage,Accession growth factor-beta (TGFB) superfamily. using the followingassays known in Tissue and Bone B02796 Bone morphogenic protein inducesbone the art: Nat Genet. 2001 January; Growth, and Diabetes formation.27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1): 295-302; J Biol Chem,Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999; and Hogan, B. L. M.(1996) Genes Dev. 10, 1580-1594. BMP-5 GeneSeq WO0020591 BMP-5 belongsto the transforming BMP-5 activity can be determined Induction ofCartilage, Accession growth factor-beta (TGFB) superfamily. using thefollowing assays known in Tissue and Bone B02797 Bone morphogenicprotein induces bone the art: Nat Genet. 2001 January; Growth, andDiabetes formation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1):295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999;and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. BMP-6 GeneSeq U.S.Pat. No. 5,187,076 BMP-6 belongs to the transforming BMP-6 activity canbe determined Induction of Cartilage, Accession growth factor-beta(TGFB) superfamily. using the following assays known in Tissue and BoneR32904 Bone morphogenic protein induces bone the art: Nat Genet. 2001January; Growth, and Diabetes formation. 27(1): 84-8; Eur J Biochem 1996Apr. 1; 237(1): 295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902,Apr. 16, 1999; and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594.Osteogenic GeneSeq WO973462 OP-1 belongs to the transforming OP-1activity can be determined Induction of Cartilage, Protein-1; OP-1;Accession growth factor-beta (TGFB) superfamily. using the followingassays known in Tissue and Bone BMP-7 W34783 Bone morphogenic proteininduces bone the art: Nat Genet. 2001 January; Growth, and Diabetesformation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1): 295-302; JBiol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999; and Hogan, B.L. M. (1996) Genes Dev. 10, 1580-1594. Osteogenic GeneSeq WO9406399 OP-2belongs to the transforming OP-2 activity can be determined Induction ofCartilage, Protein-2 Accession R57973 growth factor-beta (TGFB)superfamily. using the following assays known in Tissue and Bone Bonemorphogenic Protein induces bone the art: Nat Genet. 2001 January;Growth, and Diabetes formation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1;237(1): 295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16,1999; and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. GDP-1 GeneSeqWO9406449 Members of the TGF-beta family of The effect of GDF-1 onsignaling can Developmental Accession proteins initiate cell signalingby be assayed by treating Primary disorders, Induction of R60961 bindingto heteromeric receptor BAECs transferred with a construct Cartilage,Tissue and complexes of type I (TbetaRI) and called p3TP-Lux, containinga TGF- Bone Growth, and type II (TbetaRII) serine/threonine betaresponsive promoter fused to a Diabetes kinase receptors (reviewed byreporter gene, and measuring Massague, J. et al. (1994) Trends Cellluciferase gene expression (Wrana et Biol. 4: 172 178; Miyazono, K. etal. al., 1994, Nature 370: 341-347). (1994) Adv. Immunol. 55: 181-220).Activation of this heteromeric receptor complex occurs when TGF-betabinds to TbetaRII, which then recruits and phosphorylates TbetaRI.Activated TbetaRI then propagates the signal to downstream targets(Chen, F. and Weinberg, R. A. (1995) PNA892: 1565-1569; Wrana, J. L. etal. (1994) Nature 370: 341 347). BMP-9 GeneSeq WO9533830 BMP-9 belongsto the transforming BMP-9 activity can be determined Induction ofCartilage, Accession growth factor-beta (TGFB) superfamily. using thefollowing assays known in Tissue and Bone R86903 Bone morphogenicprotein induces bone the art: Nat Genet. 2001 January; Growth, andDiabetes formation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1):295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999;and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. BMP-10 GeneSeqWO9426893 BMP-10 belongs to the transforming BMP-10 activity can bedetermined Induction of Cartilage, Accession R66202 growth factor-beta(TGFB) superfamily. using the following assays known in Tissue and BoneBone morphogenic protein induces bone the art: Nat Genet. 2001 January;Growth, and Diabetes formation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1;237(1): 295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16,1999; and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. BMP-12GeneSeq WO9516035 BMP-12 belongs to the transforming BMP-12 activity canbe determined Induction of Cartilage, Accession R78734 growthfactor-beta (TGFB) superfamily. using the following assays known inTissue and Bone Bone morphogenic protein induces bone the art: NatGenet. 2001 January; Growth, and Diabetes formation. 27(1): 84-8; Eur JBiochem 1996 Apr. 1; 237(1): 295-302; J Biol Chem, Vol. 274, Issue 16,10897-10902, Apr. 16, 1999; and Hogan, B. L. M. (1996) Genes Dev. 10,1580-1594. BMP-15 GeneSeq W09636710 BMP-15 belongs to the transformingBMP-15 activity can be determined Induction of Cartilage, Accessiongrowth factor-beta (TGFB) superfamily. using the following assays knownin Tissue and Bone W11261 Bone morphogenic protein induces bone the art:Nat Genet. 2001 January; Growth, and Diabetes formation. 27(1): 84-8;Eur J Biochem 1996 Apr. 1; 237(1): 295-302; J Biol Chem, Vol. 274, Issue16, 10897-10902, Apr. 16, 1999; and Hogan, B. L. M. (1996) Genes Dev.10, 1580-1594. BMP-17 GeneSeq WO9929718 BMP-17 belongs to thetransforming BMP-17 activity can be determined Induction of Cartilage,Accession Y17870 growth factor-beta (TGFB) superfamily. using thefollowing assays known in Tissue and Bone Bone morphogenic proteininduces bone the art: Nat Genet. 2001 January; Growth, and Diabetesformation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1): 295-302; JBiol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999; and Hogan, B.L. M. (1996) Genes Dev. 10, 1580-1594. BMP-18 GeneSeq WO9929718 BMP-18belongs to the transforming BMP-18 activity can be determined Inductionof Cartilage, Accession Y17871 growth factor-beta (TGFB) superfamily.using the following assays known in Tissue and Bone Bone morphogenicprotein induces bone the art: Nat Genet. 2001 January; Growth, andDiabetes formation. 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1):295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999;and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. Inhibin alphaGeneSeq WO0020591 The inhibin beta A subunit joins the Tumor suppressoractivity of inhibin Tumor suppression. Accession B02806 alpha subunit toform a pituitary FSH can be determined using assays secretion inhibitor.Inhibin has been known in the art: Matzuk et al., shown to regulategonadal stromal cell Nature 1992 Nov. 26: 360 (6402); proliferationnegatively and to have 313-9. tumour-suppressor activity. In addition,serum levels of inhibin have been shown to reflect the size ofgranulosa-cell tumors and can therefore be used as a marker for primaryas well as recurrent disease. Inhibin beta GeneSeq WO0020591 The inhibinbeta A subunit joins the Tumor suppressor activity of inhibin Tumorsuppression. Accession alpha subunit to form a pituitary FSH can bedetermined using assays H02808 secretion inhibitor. Inhibin has beenknown in the art: Matzuk et al., shown to regulate gonadal stromal cellNature 1992 Nov. 26: 360 (6402); proliferation negatively and to have313-9. tumour-suppressor activity. In addition, serum levels of inhibinhave been shown to reflect the size of granulosa-cell tumors and cantherefore be used as a marker for primary as well as recurrent disease.Cerebus Protein GeneSeq WO9849296 Cerebus is believed to be involved inthe BMP activity, in the presence of the BMP Antagonist useful Accessioninhibition of BMP activity antagonist Cerebus, can be for Osteosarcoma,W86032 determined using the following abnormal bone growth. assays knownin the art: Nat Genet. 2001 January; 27(1): 84-8; Eur J Biochem 1996Apr. 1; 237(1): 295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902,Apr. 16, 1999; and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594.Soluble BMP GeneSeq WO9614579 Soluble BMP receptor kinase protein-3 BMPactivity, in the presence of the BMP Antagonist useful Receptor KinaseAccession is involved in the binding of BMPs. soluble antagonist BMPreceptor for Osteosarcoma, Protein-3 R95227 Soluble BMP receptor kinaseprotein-3 kinase protein-3, can be determined abnormal bone growth. isuseful as an antagonist for the using the following assays known ininhibition of BMP activity. the art: Nat Genet. 2001 January; 27(1):84-8; Eur J Biochem 1996 Apr. 1; 237(1): 295-302; J Biol Chem, Vol. 274,Issue 16, 10897-10902, Apr. 16, 1999; and Hogan, B. L. M. (1996) GenesDev. 10, 1580-1594. BMP Processing GeneSeq WO9741250 BMPs belong to thetransforming BMP activity, in the presence of the Bone formation orEnzyme Furin Accession growth factor-beta (TGFB) superfamily. Furin, canbe determined using the Regeneration W36099 Bone morphogenic proteininduces bone following assays known in the art: Abnormalities formation.Nat Genet. 2001 January; 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1):295-302; J Biol Chem, Vol. 274, Issue 16, 10897-10902, Apr. 16, 1999;and Hogan, B. L. M. (1996) Genes Dev. 10, 1580-1594. TGF-beta 1 GeneSeqWO9216228 Members of the TGF-beta family of The effect of TGF betas onsignaling Useful for treating Accession proteins initiate cell signalingby can be assayed by treating Primary cancer and to promote R29657binding to heteromeric receptor BAECs transfected with a construct woundhealing. complexes of type I (TbetaRI) and type called p3TP-Lux,containing a TGF- II (TbetaRII) serine/threonine kinase beta responsivepromoter fused to a receptors (reviewed by Massague, J. et reportergene, and measuring al. (1994) Trends Cell Biol. 4: 172 178; luciferasegene expression (Wrana et Miyazono, K. et al. (1994) Adv. al., 1994,Nature 370: 341-347). Immunol. 55: 181-220). Activation of thisheteromeric receptor complex occurs when TGF-beta. binds to TbetaRII,which then recruits and phosphorylates TbetaRI. Activated TbetaRI thenpropagates the signal to downstream targets (Chen, F. and Weinberg. R.A. (1995) PNA892: 1565-1569; Wrana, J. L. et al. (1994) Nature 370: 341.TGF-beta 2 GeneSeq EP542679 Members of the TGF-beta family of The effectof TGF betas on signaling Useful for treating Accession proteinsinitiate cell signaling by can be assayed by treating Primary cancer andto promote R39659 binding to heteromeric receptor BAECs transfected witha construct wound healing. complexes of type I (TbetaRI) and type calledp3TP-Lux, containing a TGF- II (TbetaRII) serine/threonine kinase betaresponsive promoter fused to a receptors (reviewed by Massague, J. etreporter gene, and measuring al. (1994) Trends Cell Biol. 4: 172 178;luciferase gene expression (Wrana et Miyazono, K. et al. (1994) Adv.al., 1994, Nature 370: 341-347). Immunol. 55: 181-220). Activation ofthis heteromeric receptor complex occurs when TGF-beta. binds toTbetaRII, which then recruits and phosphorylates TbetaRI. ActivatedTbetaRI then propagates the signal to downstream targets (Chen, F. andWeinberg. R. A. (1995) PNA892: 1565-1569; Wrana, J. L. et al. (1994)Nature 370: 341. ZTGF-beta 9 GeneSeq WO0015798 Members of the TGF-betafamily of The effect of TGF betas on signaling Useful for treatingAccession proteins initiate cell signaling by can be assayed by treatingPrimary cancer and to promote Y70654 binding to heteromeric receptorBAECs transfected with a construct wound healing. complexes of type I(TbetaRI) and type called p3TP-Lux, containing a TGF- II (TbetaRII)serine/threonine kinase beta responsive promoter fused to a receptors(reviewed by Massague, J. et reporter gene, and measuring al. (1994)Trends Cell Biol. 4: 172 178; luciferase gene expression (Wrana etMiyazono, K. et al. (1994) Adv. al., 1994, Nature 370: 341-347).Immunol. 55: 181-220). Activation of this heteromeric receptor complexoccurs when TGF-beta. binds to TbetaRII, which then recruits andphosphorylates TbetaRI. Activated TbetaRI then propagates the signal todownstream targets (Chen, F. and Weinberg. R. A. (1995) PNA892:1565-1569; Wrana, J. L. et al. (1994) Nature 370: 341. Anti-TGF betaGB2305921 Members of the TGF-beta family of The effect of TGF betas onsignaling Useful for control of family antibodies proteins initiate cellsignaling by in the presence of an anti-TGF beta fibrosis, immune, andbinding to heteromeric receptor antibody, can be assayed by treatinginflammatory disease. complexes of type I (TbetaRI) and type PrimaryBAECs transfected with a II (TbetaRII) serine/threonine kinase constructcalled p3TP-Lux, receptors (reviewed by Massague, J. et containing aTGF-beta responsive al. (1994) Trends Cell Biol. 4: 172 178; promoterfused to a reporter gene, and Miyazono, K. et al. (1994) Adv. measuringluciferase gene expression Immunol. 55: 181-220). Activation of (Wranaet al., 1994, Nature 370: 341-347). this heteromeric receptor complexoccurs when TGF-beta. binds to TbetaRII, which then recruits andphosphorylates TbetaRI. Activated TbetaRI then propagates the signal todownstream targets (Chen, F. and Weinberg. R. A. (1995) PNA892:1565-1569; Wrana, J. L. et al. (1994) Nature 370: 341. Latent TGF betaGeneSeq WO0012551 Members of the TGF-beta family of The effect of TGFbetas on signaling Useful for inhibiting binding protein II Accessionproteins initiate cell signaling by in the presence of a TGF betabinding tissue or tumor growth. Y70552 binding to heteromeric receptorprotein, can be assayed by treating complexes of type I (TbetaRI) andtype Primary BAECs transfected with a II (TbetaRII) serine/threoninekinase construct called p3TP-Lux, receptors (reviewed by Massague, J. etcontaining a TGF-beta responsive al. (1994) Trends Cell Biol. 4: 172178; promoter fused to a reporter gene, and Miyazono, K. et al. (1994)Adv. measuring luciferase gene expression Immunol. 55: 181-220).Activation of (Wrana et al., 1994, Nature 370: 341-347). thisheteromeric receptor complex occurs when TGF-beta. binds to TbetaRII,which then recruits and phosphorylates TbetaRI. Activated TbetaRI thenpropagates the signal to downstream targets (Chen, F. and Weinberg. R.A. (1995) PNA892: 1565-1569; Wrana, J. L. et al. (1994) Nature 370: 341.MP52 GeneSeq WO9741250 Members of the TGF-beta family of The effect ofTGF betas on signaling Bone formation or Accession proteins initiatecell signaling by can be assayed by treating Primary Regeneration W36100binding to heteromeric receptor BAECs transfected with a constructAbnormalities complexes of type I (TbetaRI) and type called p3TP-Lux,containing a TGF- II (TbetaRII) serine/threonine kinase beta responsivepromoter fused to a receptors (reviewed by Massague, J. et reportergene, and measuring al. (1994) Trends Cell Biol. 4: 172 178; luciferasegene expression (Wrana et Miyazono, K. et al. (1994) Adv. al., 1994,Nature 370: 341-347). Immunol. 55: 181-220). Activation of thisheteromeric receptor complex occurs when TGF-beta. binds to TbetaRII,which then recruits and phosphorylates TbetaRI. Activated TbetaRI thenpropagates the signal to downstream targets (Chen, F. and Weinberg. R.A. (1995) PNA892: 1565-1569; Wrana, J. L. et al. (1994) Nature 370: 341.b57 Protein GeneSeq WO9837195 BMPs are involved in the induction of BMPactivity, in the presence of b57 BMP Antagonist useful Accession boneformation. Specific antagonists are protein, can be determined using thefor Osteosarcoma, W69293 useful is preventing this activity fromfollowing assays known in the art: abnormal bone growth. occurring. NatGenet. 2001 January; 27(1): 84-8; Eur J Biochem 1996 Apr. 1; 237(1):295-302; J Biol Chem, Vol. 274, Issue 16, 1089-10902, Apr. 16, 1999; andHogan, B. L. M. (1996) Genes Deve. 10, 1580-1594. Resistin GeneSeqWO0064920 This gene belongs to the family defined Ability of resistin toinfluence type Type II diabetes and Accession by mouse FIZZI andFIZZ3/Resistin II diabetes can be determined using Syndrome X. W69293genes. The characteristic feature of this assays known in the art:Pontoglio et family is the C-terminal stretch of 10 cys al., J ClinInvest 1998 May 15; residues with identical spacing. The 101(10):2215-22. mouse homolog of this protein is secreted by adipocytes, may bethe hormone potantially linking obesity to type II diabetes. Galectin-4GeneSeq WO9703190 Galectins are a family of carbohydrate- Ability ofGalectin-4 polypeptides to Lactose intolerance. Accession bindingproteins characterized by an bind lactose can be determined using W11841affinity for beta-galactoside containing assays known in the art: Wada,et al., glycoconjugates. J Biol Chem 1997 Feb 28; 272(9): 6078-86.APM-I; ACRP-30; GeneSeq W00026363 ACPR30 gene is exclusively expressedin Ability of ACRP30 polypeptides to Obesity, Metabolic FamoxinAccession adipose tissue. ACRP30 is thought to influence obesity and fatoxidation disorders, Lipid Y71035 increase fatty acid oxidation bymuscle can be determined using assays Metabolism; Hormone tissue. knownin the art: Fruebis et al., Proc Secretion. Nat'l Acad Sci USA 2001 Feb.13; 98(4): 2005-10. ACRP-30 GeneSeq WO0063376 ACPR30 gene is exclusivelyexpressed in Ability of ACRP30 homologue Obesity, Metabolic Homologue;Accession adipose tissue. ACRP30 is thought to polypeptides to influenceobesity disorders, Lipid Complement B30234 increase fatty acid oxidationby muscle and fat oxidation can be determined Metabolism; HormoneComponent Clq C tissue. using assays known in the art: Secretion.Fruebis et al., Proc Nat'l Acad Sci USA 2001 Feb. 13; 98(4): 2005-10.Calpain-10a GeneSeq WO0023603 Calpain is believed to play a role inAbility of Calpain-10 to influence Diabetes mellitus; Accession insulinsecretion and insulin activity, and type II diabetes can be determinedRegulation of Insulin Y79567 therefore may be useful in the treatmentusing assays known in the art: secretory response; of type II diabetes.Pontoglio et al., J Clin Invest 1998 Insulin mediated May 15; 101(10):2215-22. glucose transport disorders. Calpain-10b GeneSeq WO0023603Calpain is believed to play a role in Ability of Calpain-10 to influenceDiabetes mellitus; Accession insulin secretion and insulin activity, andtype II diabetes can be determined Regulation of Insulin Y79568therefore may be useful in the treatment using assays known in the art:secretory response; of type II diabetes. Pontoglio et al., J Clin Invest1998 Insulin mediated May 15; 101(10): 2215-22. glucose transportdisorders. Calpain-10c GeneSeq WO0023603 Calpain is believed to play arole in Ability of Calpain-10 to influence Diabetes mellitus; Accessioninsulin secretion and insulin activity, and type II diabetes can bedetermined Regulation of Insulin Y79569 therefore may be useful in thetreatment using assays known in the art: secretory response; of type IIdiabetes. Pontoglio et al., J Clin Invest 1998 Insulin mediated May 15;101(10): 2215-22. glucose transport disorders PDGF-D GeneSeq WO0027879Vascular Endothelial Growth Factor. Proliferation assay using NR6R-3T3Wound Healing; Accession cells (Rizzino 1988 Cancer Res. 48:Atherosclermis. Y71130 4266). FasL GeneSeq WO9936079 Activitiesassociated with apoptosis and Activity can be determined usingApoptosis-related Accession immune system functions. Apoptosis assaysknown in the art: disorders; Autoimmune Y28594 Walczak et al. (1996)EMBOJ 16: disorders; Graft v-Host 5386-5397. disorders. Chondro modulin-GeneSeq W00029579 Chondromodulin proteins are cartilage Ability ofChondromodulin-like Antianglogenic agent; like protein Accessionproteins thought to confer resistance to protein to inhibitvascularization can Osteoblast proliferation Y71262 anglogeneis, andthus are useful as anti- be determined using assays known stimulator;prevents angiogenic agents that may have utility in in the art: Hirakieet al., J Biol Chem vascularization of combating cancer. 1997 Dec. 19;272(51): 32419-26. cartilage tissue; Useful to treat cancer. PatchedGeneSeq U.S. Pat. No. 5837538 Patched is a tumour-suppressor receptorAbility of soluble Patched to bind to Receptor for Hedgehog Accessionfor Sonic hedgehog (shh), which is a and inhibit the activities of shhcan cellular proliferation W72969 protein that controls developmental bedetermined using assays known signaling molecule. patterning and growth.in the art: Stone et al., Nature 1996 This receptor is useful Nov. 14;384(6605): 129-34. as a means of preventing cellular proliferation viathe shh signaling pathway, thus useful for cancers. Patched-2 GeneSeqWO9953058 Patched is a tumour-suppressor receptor Ability of solublePatched to bind to Receptor for Hedgehog Accession for Sonic hedgehog(shh), which is a and inhibit the activities of shh can cellularproliferation Y43261 protein that controls developmental be determinedusing assays known signaling molecule. patterning and growth. in theart: Stone et al., Nature 1996 This receptor is useful Nov. 14;384(6605): 129-34. as a means of preventing cellular proliferation viathe shh signaling pathway, thus useful for cancers. Maspin; ProteaseGeneSeq WO9405804 Maspin is a member of the serpin family The inhibitoryeffects of Maspin and Tumor suppressor which Inhibitor 5 Accession ofserine protease inhibitors that is other protease inhibitors can be isdown-regulated in R50938 thought to suppress tumor metastasis. assayedusing methods known in the breast cancers. The art such as a labeledprotease maspin protein has substrate, for example, Universal tumoursuppressing and Protease Substrate (casein, resorufin- invasionsuppressing labeled): Roche Molecular activity. Biochemicals, Cat. No.1080733. Endostatin GeneSeq WO0064946 Endostatin is believed to inhibiteffects of The inhibitory effects of endostatin Anti-angiogenicactivity. Accession capillary endothelial cell proliferation. can beassayed using assays Useful in the prevention B28399 disclosed by Cao etal. (1996) J. and/or treatment of Biol. Chem. 271 29461-29467. cancers.aFGF; FGF-1 GeneSeq EP298723 Fibroblast Growth Factor Proliferationassay using NR6R-3T3 Promotion of growth and Accession cells (Rizzino1988 Cancer Res. 48: proliferation of cells, P94037 4266); Examples 23and 39 such as epithelial cells disclosed herein, and keratinocytes.Antagonists may be useful as anti-cancer agents. bFGF; FGF-2 GeneSeqFR2642086 Fibroblast Growth Factor Proliferation assay using NR6R-3T3Promotion of growth and Accession cells (Rizzino 1988 Cancer Res. 48:proliferation of cells, R06685 4266); Examples 23 and 39 such asepithelial cells disclosed herein. and keratinocytes. Antagonists may beuseful as anti-cancer agents. FGF-3; INT-2 GeneSeq WO9503831 FibroblastGrowth Factor Proliferation assay using NR6R-3T3 Promotion of growth andAccession cells (Rizzino 1988 Cancer Res. 48: proliferation of cells,R07824 4266); Examples 23 and 39 such as epithelial cells disclosedherein. and keratinocytes. Antagonists may he useful as anti-canceragents. FGF-4; HST-1; GeneSeq WO9503831 Fibroblast Growth FactorProliferation assay using NR6R-3T3 Promotion of growth and HBGF-4Accession cells (Rizzino 1988 Cancer Res. 48: proliferation of cells,R07825 4266); Examples 23 and 39 such as epithelial cells disclosedherein. and keratinocytes. Antagonists may be useful as anti-canceragents. FGF-5 GeneSeq WO9730155 Fibroblast Growth Factor Proliferationassay using NR6R-3T3 Promotion of growth and Accession cells (Rizzino1988 Cancer Res. 48: proliferation of cells, W22600 4266); Examples 23and 39 such as epithelial cells disclosed herein. and keratinocytes.Antagonists may be useful as anti-cancer agents. FGF-6; Heparin GeneSeqEP613946 Fibroblast Growth Factor Proliferation assay using NR6R-3T3Promotion of growth and binding secreted Accession cells (Rizzino 1988Cancer Res. 48: proliferation of cells, transforming R58555 4266);Examples 23 and 39 such as epithelial cells factor-2 disclosed herein.and keratinocytes. Antagonists may be useful as anti-cancer agents.FGF-8 GeneSeq WO9524928 Fibroblast Growth Factor Proliferation assayusing NR6R-3T3 Promotion of growth and Accession cells (Rizzino 1988Cancer Res. 48: proliferation of cells, R80783 4266); Examples 23 and 39such as epithelial cells disclosed herein. and keratinocytes.Antagonists may be useful as anti-cancer agents. FGF-9; Gila GeneSeqWO9503831 Fibroblast Growth Factor Proliferation assay using NR6R-3T3Promotion of growth and activating factor Accession cells (Rizzino 1988Cancer Res. 48: proliferation of cells, R70822 4266); Examples 23 and 39such as epithelial cells disclosed herein. and keratinocytes.Antagonists may be useful as anti-cancer agents. FGF-12; FibroblastGeneSeq WO9635708 Fibroblast Growth Factor Proliferation assay usingNR6R-3T3 Promotion of growth and growth factor Accession cells (Rizzino1988 Cancer Res. 48: proliferation of cells, homologous W06309 4266);Examples 23 and 39 such as epithelial cells factor-1 disclosed herein.and keratinocytes. Antagonists may be useful as anti-cancer agents.FGF-15 GeneSeq WO9927100 Fibroblast Growth Factor Proliferation assayusing NR6R-3T3 Promotion of growth and Accession cells (Rizzino 1988Cancer Res. 48: proliferation of cells, Y08582 4266); Examples 23 and 39such as epithelial cells disclosed herein. and keratinocytes.Antagonists may be useful as anti-cancer agents. FGF-16 GeneSeqWO9918128 Fibroblast Growth Factor Proliferation assay using NR6R-3T3Promotion of growth and Accession cells (Rizzino 1988 Cancer Res. 48:proliferation of cells, Y05474 4266); Examples 23 and 39 such asepithelial cells disclosed herein. and keratinocytes. Antagonists may beuseful as anti-cancer agents. FGF-18 GeneSeq WO9927100 Fibroblast GrowthFactor Proliferation assay using NR6R-3T3 Promotion of growth andAccession cells (Rizzino 1988 Cancer Res. 48: proliferation of cells,Y08590 4266); Examples 23 and 39 such as epithelial cells disclosedherein. and keratinocytes. Antagonists may be useful as anti-canceragents. fit-3 ligand GeneSeq EP627487 Stem Cell Progenitor Chemokineactivities can be Promotion of immune Accession determined using assaysknown in cell growth and/or R67541 the art: Methods in Moleculardifferentiation. Biology, 2000, vol. 138: Chemokine Protocols. Editedby: A. E. I. Proudfoot, T. N. C. Wells, and C. A. Power. © Humana PressInc., Totowa,NJ VEGF-110 GeneSeq WO0013702 Promotes the growth and/orproliferation VEGF activity can be determined Promotion of growth andAccession of endothelial cells. using assays known in the art, suchproliferation of cells, Y69417 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGB-121 GeneSeq WO0071713 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, B50432 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-138 GeneSeq WO9940197 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y43483 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-145 GeneSeq WO0013702 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y69413 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-162 GeneSeq W09940197 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y43484 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-165 GeneSeq WO0013702 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y69414 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-182 GeneSeq W09940197 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y43483 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-189 GeneSeq WO0013702 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y69415 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells.. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-206 GeneSeq W00013702 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, Y69416 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-D GeneSeq WO9807832 Promotes the growthand/or proliferation VEGF activity can be determined Promotion of growthand Accession of endothelial cells. using assays known in the art, suchproliferation of cells, W53240 as those disclosed in International suchas vascular Publication No. WO0045835, for endothelial cells. example.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer. VEGF-E; VEGF-X GeneSeq W09947677 Promotes thegrowth and/or proliferation VEGF activity can be determined Promotion ofgrowth and Accession of endothelial cells. using assays known in theart, such proliferation of cells, Y33679 as those disclosed inInternational such as vascular Publication No. WO0045835, forendothelial cells. example. Antagonists may be useful as anti-angiogenicagents, and may be applicable for cancer. VEGF Receptor; GeneSeqWO9831794 Receptor for VEGF polypeptides VEGF activity, in the presenceof VEGF Receptor. Fusion KDR; flk-1 Accession flk-1 polypeptides, can beprotein with the W69679 determined using assays known in extracellulardomain is the art, such as those disclosed in useful as an anti-International Publication No. angiogenic agent. WO0045835, for example.Antagonists may be useful in the promotion of angiogenesis. Soluble VEGFGeneSeq U.S. Pat. No. 5,712,380 Receptor for VEGF polypeptides VEGFactivity, in the presence of VEGF Receptor. Fusion Receptor AccessionVEGF Receptor polypeptides, can protein with the W47037 be determinedusing assays known extracellular domain is in the art, such as thosedisclosed in useful as an anti- International Publication No. angiogenicagent. WO0045835, for example. Antagonists may be useful in thepromotion of angiogenesis. flt-1 GeneSeq WO0021560 Receptor for VEGFpolypeptides VEGF activity, in the presence of VEGF Receptor. FusionAccession flt-1 polypeptides, can be protein with the Y70751 determinedusing assays known in extracellular domain is the art, such as thosedisclosed in useful as an anti- International Publication No. angiogenicagent. WO0045835, for example. Antagonists may be useful in thepromotion of angiogenesis. VEGF R-3; flt-4 GeneSeq WO0058511 Receptorfor VEGF polypeptides VEGF activity, in the presence of VEGF Receptor.Fusion Accession flt-4 polypeptides, can be protein with the B29047determined using assays known in extracellular domain is the art, suchas those disclosed in useful as an anti- International Publication No.angiogenic agent. WO0045835, for example. Antagonists may be useful inthe promotion of angiogenesis. Neuropilin-1 GeneSeq WO9929858 VascularEndothelial Growth Factor VEGF activity can be determined Promotion ofgrowth and Accession using assays known in the art, such proliferationof cells, Y06319 as those disclosed in International such as vascularPublication No. WO0045835, for endothelial cells. example. Antagonistsmay be useful as anti-angiogenic agents, and may be applicable forcancer. Neuropilin-2 GeneSeq WO9929858 Vascular Endothelial GrowthFactor VEGF activity can be determined Promotion of growth and Accessionusing assays known in the art, such proliferation of cells, Y03618 asthose disclosed in International such as vascular Publication No.WO0045835, for endothelial cells. example. Antagonists may be useful asanti-angiogenic agents, and may be applicable for cancer. Human fasttwitch GeneSeq W09730085 Troponins are contractile proteins that areAbility of soluble Troponins to Anti-angiogenesis skeletal muscleAccession thought to inhibit angiogenesis. High inhibit anglogenesis canbe troponin C W22597 levels may contribute to the difficulty determinedusing assays known in encountered in revascularizing the the art:. ProcNatl Acad Sci USA ischemic myocardium after 1999 Mar. 16; 96(6):2645-50. cardiovascular injury. Human fast twitch GeneSeq W09730085Troponins are contractile proteins that are Ability of soluble Troponinsto Anti-angiogenesis skeletal muscle Accession thought to inhibitangiogenesis. High inhibit anglogenesis can be troponin I W18054 levelsmay contribute to the difficulty determined using assays known inencountered in revascularizing the the art:. Proc Natl Acad Sci USAischemic myocardium after 1999 Mar. 16; 96(6): 2645-50. cardiovascularinjury. Human fast twitch GeneSeq W09730085 Troponins are contractileproteins that are Ability of soluble Troponins to Anti-angiogenesisskeletal muscle Accession thought to inhibit angiogenesis. High inhibitanglogenesis can be troponin T W22599 levels may contribute to thedifficulty determined using assays known in encountered inrevascularizing the the art:. Proc Natl Acad Sci USA ischemic myocardiumafter 1999 Mar. 16; 96(6): 2645-50. cardiovascular injury. fragment.GeneSeq W09719955 Troponins are contractile proteins that are Ability ofsoluble Troponins to Anti-angiogenesis myofibrillar Accession thought toinhibit angiogenesis. High inhibit anglogenesis can be protein troponinI W18053 levels may contribute to the difficulty determined using assaysknown in encountered in revascularizing the the art:. Proc Natl Acad SciUSA ischemic myocardium after 1999 Mar. 16; 96(6): 2645-50.cardiovascular injury. myofibrillar GeneSeq W09719955 Troponins arecontractile proteins that are Ability of soluble Troponins toAnti-angiogenesis protein troponin I Accession thought to inhibitangiogencsis. High inhibit anglogenesis can be W18054 levels maycontribute to the difficulty determined using assays known inencountered in revascularizing the the art:. Proc Natl Acad Sci USAischemic myocardium after 1999 Mar. 16; 96(6): 2645-50. cardiovascularinjury. Troponin peptides GeneSeq WO9933874 Troponins are contractileproteins that are Ability of soluble Troponins to Anti-angiogenesisAccessions thought to inhibit angiogencsis. High inhibit anglogenesiscan be Y29581, Y29582, levels may contribute to the difficultydetermined using assays known in Y29583, Y29584, encountered inrevascularizing the the art:. Proc Natl Acad Sci USA Y29585, andischemic myocardium after 1999 Mar. 16; 96(6): 2645-50. Y29586cardiovascular injury. Human fast twitch GeneSeq WO0054770 Troponins arecontractile proteins that are Ability of soluble Troponins toAnti-angiogenesis skeletal muscle Accession thought to inhibitangiogencsis. High inhibit anglogenesis can be Troponin subunit C B00134levels may contribute to the difficulty determined using assays known inencountered in revascularizing the the art:. Proc Natl Acad Sci USAischemic myocardium after 1999 Mar. 16; 96(6): 2645-50. cardiovascularinjury. Human fast twitch GeneSeq WO0054770 Troponins are contractileproteins that are Ability of soluble Troponins to Anti-angiogenesisskeletal muscle Accession thought to inhibit angiogencsis. High inhibitanglogenesis can be Troponin subunit I B00135 levels may contribute tothe difficulty determined using assays known in Protein encountered inrevascularizing the the art:. Proc Natl Acad Sci USA ischemic myocardiumafter 1999 Mar. 16; 96(6): 2645-50. cardiovascular injury. Human fasttwitch GeneSeq WO0054770 Troponins are contractile proteins that areAbility of soluble Troponins to Anti-angiogenesis skeletal muscleAccession thought to inhibit angiogencsis. High inhibit anglogenesis canbe Troponin subunit T B00136 levels may contribute to the difficultydetermined using assays known in encountered in revascularizing the theart:. Proc Natl Acad Sci USA ischemic myocardium after 1999 Mar. 16;96(6): 2645-50. cardiovascular injury. Activator GeneSeq WO9013648 PAIsare believed to play a role in cancer, Methods that measure plasminogenAnti-angiogenesis; Inbibitor-1; PAI-1 Accession and cardiovasculardisease and blood- activator inhibitor (PAI) activity are blood-clottingdisorders. R08411 clotting disorders. known in the art, for example,assay the ability of PAI to inhibit tissue plasminogen activator (tPA)or urokinase (uPA): J Biochem Biophys Methods 2000 Sep. 11; 45(2):127-40, Breast Cancer Res Treat 1996; 41(2): 141-6. Methods that measureanti-angiogenesis activity are known in the art, for example, Proc NatlAcad Sci USA 1999 Mar. 16; 96(6): 2645-50. Plasminogen GeneSeq DE3722673PAIs are believed to play a role in cancer, Methods that measureplasminogen Anti-angiogenesis; Activator Accession and cardiovasculardisease and blood- activator inhibitor (PAI) activity are blood-clottingdisorders. Inhibitor-2; PAI-2 P94160 clotting disorders. known in theart, for example, assay the ability of PAI to inhibit tissue plasminogenactivator (tPA) or urokinase (uPA): J Biochem Biophys Methods 2000 Sep.11; 45(2): 127-40, Breast Cancer Res Treat 1996; 41(2): 141-6. Methodsthat measure anti-angiogenesis activity are known in the art, forexample, Proc Natl Acad Sci USA 1999 Mar. 16; 96(6): 2645-50. ActivatorGeneSeq WO9102057 PAIs are believed to play a role in cancer, Methodsthat measure plasminogen Anti-angiogenesis; Inhibitor-2; PAI-2 Accessionand cardiovascular disease and blood- activator inhibitor (PAI) activityare blood-clotting disorders. R10921 clotting disorders. known in theart, for example, assay the ability of PAI to inhibit tissue plasminogenactivator (tPA) or urokinase (uPA): J Biochem Biophys Methods 2000 Sep.11; 45(2): 127-40, Breast Cancer Res Treat 1996; 41(2): 141-6. Methodsthat measure anti-angiogenesis activity are known in the art, forexample, Proc Natl Acad Sci USA 1999 Mar. 16; 96(6): 2645-50. HumanPAI-1 GeneSeq WO9105048 PAIs are believed to play a role in cancer,Methods that measure plasminogen Anti-angiogenesis; mutants Accessionsand cardiovascular disease and blood- activator inhibitor (PAI) activityare blood-clotting disorders. R11755, R11756, clotting disorders. knownin the art, for example, assay R11757, R11758, the ability of PAI toinhibit tissue R11759, R11760, plasminogen activator (tPA) or R11761,R11762 urokinase (uPA): J Biochem and R11763 Biophys Methods 2000 Sep.11; 45(2): 127-40, Breast Cancer Res Treat 1996; 41(2): 141-6. Methodsthat measure anti-angiogenesis activity are known in the art, forexample, Proc Natl Acad Sci USA 1999 Mar. 16; 96(6): 2645-50. CXCR3; CXCGeneSeq WO0018431 Chemokines are a family of related small, Chemokineactivities can be Soluble CXCR3 Accession secreted proteins involved inbiological determined using assays known in polypeptides may be Y79372processes ranging from hematopoiesis, the art: Methods in Molecularuseful for inhibiting angiogenesis, and leukocyte trafficking. Biology,2000, vol. 138: Chemokine chemokine activities and Members of thisfamily are involved in a Protocols. Edited by: A. E. I. Proudfoot, viralinfection. similarly diverse range of pathologies T. N. C. Wells, and C.A. Power. including inflammation, allergy, tissue © Humana Press Inc.,rejection, viral infection, and tumor Totowa, NJ. biology. Thechemokines exert their effects by acting on a family of seventransmembrane G-protein coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified.Modified Rantes GeneSeq WO9737005 Chemokines are a family of relatedsmall, Chemokine activities can be Immune disorders. Accession secretedproteins involved in biological determined using assays known in W38129processes ranging from hematopoiesis, the art: Methods in Molecularangiogenesis, and leukocyte trafficking. Biology, 2000, vol. 138:Chemokine Members of this family are involved in a Protocols. Edited by:A. E. I. Proudfoot, similarly diverse range of pathologies T. N. C.Wells, and C. A. Power. including inflammation, allergy, tissue © HumanaPress Inc., rejection, viral infection, and tumor Totowa, NJ. biology.The chemokines exert their effects by acting on a family of seventransmembrane G-protein coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. RANTESGeneSeq EP905240 Chemokines are a family of related small, Chemokineactivities can be Immune disorders. Accession secreted proteins involvedin biological determined using assays known in Y05299 processes rangingfrom hematopoiesis, the art: Methods in Molecular angiogenesis, andleukocyte trafficking. Biology, 2000, vol. 138: Chemokine Members ofthis family are involved in a Protocols. Edited by: A. E. I. Proudfoot,similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-proteincoupled receptors. Over 40 human chemokines have been described, whichbind to ~17 receptors thus far identified. MCl-la GeneSeq WO9509232Chemokines are a family of related small, Chemokine activities can beImmune disorders. Accession secreted proteins involved in biologicaldetermined using assays known in R73914 processes ranging fromhematopoiesis, the art: Methods in Molecular angiogenesis; and leukocytetrafficking. Biology, 2000, vol. 138: Chemokine Members of this familyare involved in a Protocols. Edited by: A. E. I. Proudfoot, similarlydiverse range of pathologies T. N. C. Wells, and C. A. Power. includinginflammation, allergy, tissue © Humana Press Inc., rejection, viralinfection, and tumor Totowa, NJ. biology. The chemokines exert theireffects by acting on a family of seven transmembrane G-protein coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. MCP-lb GeneSeq WO9929728 Chemokinesare a family of related small, Chemokine activities can be Immunedisorders. Accession secreted proteins involved in biological determinedusing assays known in Y26176 processes ranging from hematopoiesis, theart: Methods in Molecular angiogenesis, and leukocyte trafficking.Biology, 2000, vol. 138: Chemokine Members of this family are involvedin a Protocols. Edited by: A. E. I. Proudfoot, similarly diverse rangeof pathologies T. N. C. Wells, and C. A. Power. including inflammation,allergy, tissue © Humana Press Inc., rejection, viral infection, andtumor Totowa, NJ. biology. The chemokines exert their effects by actingon a family of seven transmembrane G-protein coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. MCP-1 receptor GeneSeq WO9519436 Chemokines are a familyof related small, Chemokine activities can be Soluble MCP-1 ReceptorAccession secreted proteins involved in biological determined usingassays known in polypeptides may be R79165 processes ranging fromhematopoiesis, the art: Methods in Molecular useful for inhibitingangiogenesis, and leukocyte trafficking. Biology, 2000, vol. 138:Chemokine chemokine activities and Members of this family are involvedin a Protocols. Edited by: A. E. I. Proudfoot, viral infection.similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-proteincoupled receptors. Over 40 human chemokines have been described, whichbind to ~17 receptors thus far identified. MCP-3 GeneSeq W09509232Chemokines are a family of related small, Chemokine activities can beImmune disorders. Accession secreted proteins involved in biologicaldetermined using assays known in R73915 processes ranging fromhematopoiesis, the art: Methods in Molecular angiogenesis, and leukocytetrafficking. Biology, 2000, vol. 138: Chemokine Members of this familyare involved in a Protocols. Edited by: A. E. I. Proudfoot, similarlydiverse range of pathologies T. N. C. Wells, and C. A. Power. includinginflammation, allergy, tissue © Humana Press Inc., rejection, viralinfection, and tumor Totowa, NJ. biology. The chemokines exert theireffects by acting on a family of seven transmembrane G-protein coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. MCP-4 receptor GeneSeq W09809171Chemokines are a family of related small, Chemokine activities can beSoluble MCP-4 Receptor Accession secreted proteins involved inbiological determined using assays known in polypeptides may be W56689processes ranging from hematopoiesis, the art: Methods in Molecularuseful for inhibiting angiogenesis, and leukocyte trafficking. Biology,2000, vol. 138: Chemokine chemokine activities and Members of thisfamily are involved in a Protocols. Edited by: A. E. I. Proudfoot, viralinfection. similarly diverse range of pathologies T. N. C. Wells, and C.A. Power. including inflammation, allergy, tissue © Humana Press Inc.,rejection, viral infection, and tumor Totowa, NJ. biology. Thechemokines exert their effects by acting on a family of seventransmembrane G-protein coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. RANTESreceptor GeneSeq U.S. Pat. No. 5,652,133 Chemokines are a family ofrelated small, Chemokine activities can be Soluble RANTES Accessionsecreted proteins involved in biological determined using assays knownin Receptor polypeptides W29588 processes ranging from hematopoiesis,the art: Methods in Molecular may be useful for angiogenesis, andleukocyte trafficking. Biology, 2000, vol. 138: Chemokine inhibitingchemokine Members of this family are involved in a Protocols. Edited by:A. E. I. Proudfoot, activities and viral similarly diverse range ofpathologies T. N. C. Wells, and C. A. Power. infection. includinginflammation, allergy, tissue © Humana Press Inc., rejection, viralinfection, and tumor Totowa, NJ. biology. The chemokines exert theireffects by acting on a family of seven transmembrane G-protein coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. CCR5 variant GeneSeq WO9854317Chemokines are a family of related small, Chemokine activities can beSoluble CCR5 Accession secreted proteins involved in biologicaldetermined using assays known in polypeptides may be W88238 processesranging from hematopoiesis, the art: Methods in Molecular useful forinhibiting angiogenesis, and leukocyte trafficking. Biology, 2000, vol.138: Chemokine chemokine activities and Members of this family areinvolved in a Protocols. Edited by: A. E. I. Proudfoot, viral infection.similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-proteincoupled receptors. Over 40 human chemokines have been described, whichbind to ~17 receptors thus far identified. CCR7 GeneSeq U.S. Pat. No.6,153,441 Chemokines are a family of related small, Chemokine activitiescan be Soluble CCR7 Accession secreted proteins involved in biologicaldetermined using assays known in polypeptides may be B50859 processesranging from hematopoiesis, the art: Methods in Molecular useful forinhibiting angiogenesis, and leukocyte trafficking. Biology, 2000, vol.138: Chemokine chemokine activities and Members of this family areinvolved in a Protocols. Edited by: A. E. I. Proudfoot, viral infection.similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-proteincoupled receptors. Over 40 human chemokines have been described, whichbind to ~17 receptors thus far identified. CXC3 GeneSeq WO9727299Chemokines are a family of related small, Chemokine activities can beImmune disorders. Accession secreted proteins involved in biologicaldetermined using assays known in W23345 processes ranging fromhematopoiesis, the art: Methods in Molecular angiogenesis, and leukocytetrafficking. Biology, 2000, vol. 138: Chemokine Members of this familyare involved in a Protocols. Edited by: A. E. I. Proudfoot, similarlydiverse range of pathologies T. N. C. Wells, and C. A. Power. includinginflammation, allergy, tissue © Humana Press Inc., rejection, viralinfection, and tumor Totowa, NJ. biology. The chemokines exert theireffects by acting on a family of seven transmembrane G-protein coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Eotaxin GeneSeq WO9700960 Chemokinesare a family of related small, Chemokine activities can be Immunedisorders. Accession secreted proteins involved in biological determinedusing assays known in W10099 processes ranging from hematopoiesis, theart: Methods in Molecular angiogenesis, and leukocyte trafficking.Biology, 2000, vol. 138: Chemokine Members of this family are involvedin a Protocols. Edited by: A. E. I. Proudfoot, similarly diverse rangeof pathologies T. N. C. Wells, and C. A. Power. including inflammation,allergy, tissue © Humana Press Inc., rejection, viral infection, andtumor Totowa, NJ. biology. The chemokines exert their effects by actingon a family of seven transmembrane G-protein coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Neurotactin GeneSeq U.S. Pat. No. 6,013,257 Neurotactinmay play a role in Chemotactic leukocyte migration Immune disorders.Accessions WO9742224 chemotactic leukocyte migration and assays areknown in the art, for Y77537, W34307, brain inflammation processes.example: J. Immunol. Methods 33, Y53259, and, ((1980)); Nature 1997 Jun5; Y77539 387(6633): 611-7. Human CKbeta-9 GeneSeq U.S. Pat. No.6,153,441 Chemokines are a family of related small, Chemokine activitiescan be Immune disorders. Accession secreted proteins involved inbiological determined using assays known in B50860 processes rangingfrom hematopoiesis, the art: Methods in Molecular angiogenesis, andleukocyte trafficking. Biology, 2000, vol. 138: Chemokine Members ofthis family are involved in a Protocols. Edited by: A. E. I. Proudfoot,similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-proteincoupled receptors. Over 40 human chemokines have been described, whichbind to ~17 receptors thus far identified. Lymphotactin GeneSeqWO0073320 Chemokines are a family of related small, Chemokine activitiescan be Immune disorders. Accession secreted proteins involved inbiological determined using assays known in B50052 processes rangingfrom hematopoiesis, the art: Methods in Molecular angiogenesis, andleukocyte trafficking. Biology, 2000, vol. 138: Chemokine Members ofthis family are involved in a Protocols. Edited by: A. E. I. Proudfoot,similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G. MIP-3alpha GeneSeq WO9801557 Chemokines are a family of related small,Chemokine activities can be Immune disorders. Accession secretedproteins involved in biological determined using assays known in W44398processes ranging from hematopoiesis, the art: Methods in Molecularangiogenesis, and leukocyte trafficking. Biology, 2000, vol. 138:Chemokine Members of this family are involved in a Protocols. Edited by:A. E. I. Proudfoot, similarly diverse range of pathologies T. N. C.Wells, and C. A. Power. including inflammation, allergy, tissue © HumanaPress Inc., rejection, viral infection, and tumor Totowa, NJ. biology.The chemokines exert their effects by acting on a family of seventransmembrane G. MIP-3 beta GeneSeq WO9801557 Chemokines are a family ofrelated small, Chemokine activities can be Immune disorders. Accessionsecreted proteins involved in biological determined using assays knownin W44399 processes ranging from hematopoiesis, the art: Methods inMolecular angiogenesis, and leukocyte trafficking. Biology, 2000, vol.138: Chemokine Members of this family are involved in a Protocols.Edited by: A. E. I. Proudfoot, similarly diverse range of pathologies T.N. C. Wells, and C. A. Power. including inflammation, allergy, tissue© Humana Press Inc., rejection, viral infection, and tumor Totowa, NJ.biology. The chemokines exert their effects by acting on a family ofseven transmembrane G. MIP-Gamma GeneSeq WO9504158 Chemokines are afamily of related small, Chemokine activities can be Immune disorders.Accession secreted proteins involved in biological determined usingassays known in R70798 processes ranging from hematopoiesis, the art:Methods in Molecular angiogenesis, and leukocyte trafficking. Biology,2000, vol. 138: Chemokine Members of this family are involved in aProtocols. Edited by: A. E. I. Proudfoot, similarly diverse range ofpathologies T. N. C. Wells, and C. A. Power. including inflammation,allergy, tissue © Humana Press Inc., rejection, viral infection, andtumor Totowa, NJ. biology. The chemokines exert their effects by actingon a family of seven transmembrane G. Stem Cell GeneSeq WO9104274Chemokines are a family of related small, Chemokine activities can beHematopoietic growth Inhibitory Accession secreted proteins involved inbiological determined using assays known in factors. Factor R11553processes ranging from hematopoiesis, the art: Methods in Molecularangiogenesis, and leukocyte trafficking. Biology, 2000, vol. 138:Chemokine Members of this family are involved in a Protocols. Edited by:A. E. I. Proudfoot, similarly diverse range of pathologies T. N. C.Wells, and C. A. Power. including inflammation, allergy, tissue © HumanaPress Inc., rejection, viral infection, and tumor Totowa, NJ. biology.The chemokines exert their effects by acting on a family of seventransmembrane G. thrombopoietin GeneSeq WO9521920 Thrombopoietin isinvolved in the Thrombopoietin (TPO) can be Hematopoietic growthAccession regulation of the growth and assayed to determine regulationof factors. R79905 differentiation of megakaryocytes and growth anddifferentiation of preceptors thereof. megakaryocytes. Mol Cell Biol2001 April; 21(8): 2659-70; Exp Hematol 2001 January; 29(1): 51-8 andwithin. c-kit ligand; GeneSeq EP992579 and C-kit ligan is thought tostimulate the Chemokine activities can be Hematopoietic growth SCF; Mastcell Accession EP676470 proliferation of mast cells, and is able todetermined using assays known in factors. growth factor; Y53284, R83978augment the proliferation of both the art: Methods in Molecular MGF; andR83977 myeloid and lymphoid hematopoietic Biology, 2000, vol. 138:Chemokine Fibrosarcoma- progenitors in bone marrow culture. C-Protocols. Edited by: A. E. I. Proudfoot, derived stem kit ligand isalso though to act T. N. C. Wells, and C. A. Power. cell factorsynergistically with other cytokines. © Humana Press Inc., Totowa, NJ.Platelet derived GeneSeq WO0066736 Vascular Endothelial Growth FactorVEGF activity can be determined Promotion of growth and growth factorAccession B48653 using assays known in the art, such proliferation ofcells, as those disclosed in International such as vascular PublicationNo. WO0045835, for endothelial cells. example. Antagonists may be usefulas anti-angiogenic agents, and may be applicable for cancer. MelanomaGeneSeq WO9503328 Melanoma inhibiting protein has Tumor suppressoractivity of Cancer; melanoma inhibiting protein Accession R69811melanoma-inhibiting activity and can be melanoma inhibiting protein canbe used to treat cancer (melanoma, determined using assays known inglioblastoma, neuroblastoma, small cell the art: Matzuk et al., Nature1992 lung cancer, neuroectodermal tumors) or Nov 26; 360(6402): 313-9.as an immunosuppressant (it inhibits IL-2 or phytohaemagglutinin inducedproliferation of peripheral blood lymphocytes. Glioma-derived GeneSeqEP399816 Vascular Endothelial Growth Factor VEGF activity can bedetermined Promotion of growth and growth factor Accession R08120 usingassays known in the art, such proliferation of cells, as those disclosedin International such as vascular Publication No. WO0045835, forendothelial cells. example. Antagonists may be useful as anti-angiogenicagents, and may be applicable for cancer. Platelet derived GeneSeqEP682110 Vascular Endothelial Growth Factor VEGF activity can bedetermined Promotion of growth and growth factor Accession R84759 usingassays known in the art, such proliferation of cells, precursor A asthose disclosed in International such as vascular Publication No.WO0045835, for endothelial cells. example. Antagonists may be useful asanti-angiogenic agents, and may be applicable for cancer. Plateletderived GeneSeq EP682110 Vascular Endothelial Growth Factor VEGFactivity can be determined Promotion of growth and growth factorAccession R84760 using assays known in the art, such proliferation ofcells, precursor B as those disclosed in International such as vascularPublication No. WO0045835, for endothelial cells. example. Antagonistsmay be useful as anti-angiogenic agents, and may be applicable forcancer. Platelet derived GeneSeq EP282317 Vascular Endothelial GrowthFactor VEGF activity can be determined Promotion of growth and growthfactor Bv- Accession P80595 using assays known in the art, suchproliferation of cells, sis and P80596 as those disclosed inInternational such as vascular Publication No. WO0045835, forendothelial cells. example. Antagonists may be useful as anti-angiogenicagents, and may be applicable for cancer. Placental Growth GeneSeqWO9206194 Vascular Endothelial Growth Factor VEGF activity can bedetermined Promotion of growth and Factor Accessions using assays knownin the art, such proliferation of cells, R23059 and as those disclosedin International such as vascular R23060 Publication No. WO0045835, forendothelial cells. example. Antagonists may be useful as anti-angiogenicagents, and may be applicable for cancer. Placental Growth GeneSeqDE19748734 Vascular Endothelial Growth Factor VEGF activity can bedetermined Promotion of growth and Factor-2 Accession Y08289 usingassays known in the art, such proliferation of cells, as those disclosedin International such as vascular Publication No. WO0045835, forendothelial cells. example. Antagonists may be useful as anti-angiogenicagents, and may be applicable for cancer. Thrombopoietin GeneSeqWO0000612 Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative1 Accession Y77244 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Thrombopoietin GeneSeq WO0000612Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative2 Accession Y77255 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Thrombopoietin GeneSeq WO0000612Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative3 Accession Y77262 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Thrombopoietin GeneSeq WO0000612Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative4 Accession Y77267 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Thrombopoietin GeneSeq WO0000612Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative5 Accession Y77246 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Thrombopoietin GeneSeq WO0000612Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative6 Accession Y77253 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Thrombopoietin GeneSeq WO0000612Thrombopoietin is involved in the Thrombopoietin (TPO) can beThrombocytopenia, derivative7 Accession Y77256 regulation of the growthand assayed to determine regulation of cancer. differentiation ofmegakaryocytes and growth and differentiation of preceptors thereof.megakaryocytes. Mol Cell Biol 2001 April; 21(8): 2659-70; Exp Hematol2001 January; 29(1): 51-8 and within. Fractalkine GeneSeq U.S. Pat. No.6,043,086 Fractalkine is believed to play a role in Fractalkine activitycan be Immune disorders. Accession Y53255 chemotactic leukocytemigration and determined using Chemotactic neurological disorders.leukocyte migration assays known in the art, for example: J. Immunol.Methods 33, ((1980)); Nature 1997 Jun 5; 387(6633): 611-7. CXC3 GeneSeqWO9757599 Chemokines are a family of related small, Chemokine activitiescan be Immune disorders. Accession secreted proteins involved inbiological determined using assays known in W23345 processes rangingfrom hematopoiesis, the art: Methods in Molecular angiogenesis, andleukocyte trafficking. Biology, 2000, vol. 138: Chemokine Members ofthis family are involved in a Prototols. Edited by: A. E. I. Proudfoot,similarly diverse range of pathologies T. N. C. Wells, and C. A. Power.including inflammation, allergy, tissue © Humana Press Inc., rejection,viral infection, and tumor Totowa, NJ. biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-proteincoupled receptors. Over 40 human chemokines have been described, whichbind to ~17 receptors thus far identified. CCR7 GeneSeq U.S. Pat. No.6,153,441 Chemokines are a family of related small, Chemokine activitiescan be Soluble CCR7 Accession B50859 secreted proteins involved inbiological determined using assays known in polypeptides may beprocesses ranging from hematopoiesis, the art: Methods in Molecularuseful for inhibiting angiogenesis, and leukocyte trafficking. Biology,2000, vol. 138: Chemokine chemokine activities and Members of thisfamily are involved in a Protocols. Edited by: A. E. I. Proudfoot, viralinfection. similarly diverse range of pathologies T. N. C. Wells, and C.A. Power. including inflammation, allergy, tissue © Humana Press Inc.,rejection, viral infection, and tumor Totowa, NJ. biology. Thechemokines exert their effects by acting on a family of seventransmembrane G-protein coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. NerveGrowth GeneSeq EP414151 Nerve Growth Factor Proliferation assay usingNR6R-3T3 Neurological disorders, Factor-beta Accession R11474 cells(Rizzino 1988 Cancer Res. 48: cancer 4266) Nerve Growth GeneSeq EP859056Nerve Growth Factor Proliferation assay using NR6R 3T3 Neurologicaldisorders, Factor-beta2 Accession cells (Rizzino 1988 Cancer Res. 48:cancer W69725 4266 Neurotrophin-3 GeneSeq WO9821234 Neurotrophinsregulate neuronal cell Trk tyrosine kinase activation assaysNeurological disorders, Accession survival and synaptic plasticity.known in the art can be used to assay cancer W8889 for neurotrophinactivity, for example, Proc Natl Acad Sci USA 2001 Mar 13; 98(6):3555-3560. Neurotrophin-3 GeneSeq WO9325684 Neurotrophins regulateneuronal cell Trk tyrosine kinase activation assays Neurologicaldisorders, Accession R47100 survival and synaptic plasticity. known inthe art can be used to assay cancer for neurotrophin activity, forexample, Proc Natl Acad Sci USA 2001 Mar 13; 98(6): 3555-3560.Neurotrophin-4a GeneSeq WO9325684 Neurotrophins regulate neuronal cellTrk tyrosine kinase activation assays Neurological disorders, AccessionR47101 survival and synaptic plasticity. known in the art can be used toassay cancer for neurotrophin activity, for example, Proc Natl Acad SciUSA 2001 Mar 13; 98(6): 3555-3560. 13; 98(6): 3555-3560 Neurotrophin-4bGeneSeq WO9325684 Neurotrophins regulate neuronal cell Trk tyrosinekinase activation assays Neurological disorders, Accession R47102survival and synaptic plasticity. known in the art can be used to assaycancer tyrosine kinases. for neurotrophin activity, for example, ProcNatl Acad Sci USA 2001 Mar 13; 98(6): 3555-3560. Neurotrophin-4c GeneSeqWO9325684 Neurotrophins regulate neuronal cell Trk tyrosine kinaseactivation assays Neurological disorders, Accession R47103 survival andsynaptic plasticity. known in the art can be used to assay cancertyrosine kinases. for neurotrophin activity, for example, Proc Natl AcadSci USA 2001 Mar 13; 98(6): 3555-3560. Neurotrophin-4d GeneSeq WO9325684Neurotrophins regulate neuronal cell Trk tyrosine kinase activationassays Neurological disorders, Accession R47102 survival and synapticplasticity. known in the art can be used to assay cancer tyrosinekinases. for neurotrophin activity, for example, Proc Natl Acad Sci USA2001 Mar 13; 98(6): 3555-3560. Platelet-Derived GeneSeq U.S. Pat. No.5,219,739 Vascular Endothelial Growth Factor VEGF activity can bedetermined Promotion of growth and Growth Factor Accession R38918 usingassays known in the art, such as proliferation of cells, A chain thosedisclosed in International such as vascular Publication No. W00045835,for endothelial cells. example. Hematopoietic and immune disorders.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer Platelet-Derived GeneSeq U.S. Pat. No. 5,219,739Vascular Endothelial Growth Factor VEGF activity can be determinedPromotion of growth and Growth Factor Accession R38919 using assaysknown in the art, such as proliferation of cells, B chain thosedisclosed in International such as vascular Publication No. W00045835,for endothelial cells. example. Hematopoietic and immune disorders.Antagonists may be useful as anti-angiogenic agents, and may beapplicable for cancer Stromal Derived GeneSeq WO9948528 Stromal GrowthFactor Proliferation assay using NR6R-3T3 Hematopoietic, immune Factor-1alpha Accession cells (Rizzino 1988 Cancer Res. 48: disorders, cancerY39995 4266) Stromal Derived GeneSeq CA2117953 Stromal Growth FactorProliferation assay using NR6R-3T3 Hematopoietic, immune Factor-1 betaAccession cells (Rizzino 1988 Cancer Res. 48: disorders, cancer R754204266) Tarc GeneSeq WO9711969 Chemotactic for T lymphocytes. MayChemotactic leukocyte migration Antiinflammatory. Accession play a rolein T-cell development. assays are known in the art, for Immunedisorders, W14917 Thought to bind CCR8 and CCR4 example: J. Immunol.Methods 33 cancer ((1980)) Prolactin GeneSeq WO9521625 Prolactin isinvolved in immune cell Immune coil proliferation and Reproductivesystem Accession R78691 proliferation and apoptosis. suppression ofapoptosis by prolactin disorders, cancer. can be assayed by methodswell- known in the art, for example, Buckley, AR and Buckley DJ, Ann N YAcad Sci 2000; 917: 522-33, and within. Prolactin2 GeneSeq U.S. Pat. No.5,955,346 Prolactin is involved in immune cell Immune coil proliferationand Reproductive system Accession proliferation and apoptosis.suppression of apoptosis by prolactin disorders, cancer. Y31764 can beassayed by methods well- known in the art, for example, Buckley, AR andBuckley DJ, Ann N Y Acad Sci 2000; 917: 522-33, and within. FollicleGeneSeq EP974359 FSH stimulates secretion of interleukin-1 FSHactivities can be determined Reproductive system stimulating Accessionby cells isolated from women in the using assays known in the art; JGend disorders, cancer. hormone Alpha Y54160 follicular phase Specif Med1999 November-December; 2(6): 30-4; subunit Mol Cell Endocrinol. 1997Nov 15; 134(2): 109-18. Follicle GeneSeq EP974359 FSH stimulatessecretion of interleukin-1 FSH activities can be determined Reproductivesystem stimulating Accession by cells isolated from women in the usingassays known in the art; J Gend disorders, cancer. hormone Beta Y54161follicular phase Specif Med 1999 November-December; 2(6): 30-4; subunitMol Cell Endocrinol. 1997 Nov 15; 134(2): 109-18. Substance P GeneSeqWO0054053 Substance P is associated with Immuneregulation and bonemarrow, diabetes mellitus, (tachykinin) Accession immunoregulation. cellproliferation by substance P can hypertension, cancer B23027 be assayedby methods well-known in the art, for example, Lai et al. Proc Natl AcadSci USA 2001 Mar 27; 98(7): 3970-5; Jallat-Daloz et al. Allergy AsthmaProc 2001 January-February; 22(1): 17-23; Kahler et al. Exp Lung Res2001 January-February; 27(1): 25-46; and Adamus MA and Dabrowski ZJ. JCell Biochem 2001; 81(3)499-506. Ocytocin GeneSeq WO0053755 Oxytocin isinvolved in the induction of Oxytocin and prostaglandin E(2)inflammatory disorders (Neurophysin I) Accession prostaglandin (E2)release as well as an release and Ocytocin (Ca2+) increase immunologicdisorders, B24085 and increased amount of calcium release by can beassayed by methods well- cancer B24086 smooth muscle cells. known in theart, for example, Pavan et al., AM J Obset Gynecol 2000 July; 183(1):76-82 and Holda et al., Cell Calcium 1996 July; 20(1): 43 51.Vasopressin GeneSeq WO0053755 Vasopressinis believed to have a directVasopressin activity can be inflammatory disorders (Neurophysin II)Accession antidiuretic action on the kidney, and it is determined usingassays known in the immunologic disorders, B24085 and thought to causevasoconstriction of the art, for example, Endocr Regul 1996 cancerB24086 peripheral vessels. March; 30(1): 13-17. IL-1 GeneSeq EP165654Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, P60326lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Orencole &Dinarclio (1989) neutrophils and T lymphocytes, and/or Cytokine 1,14-20. inhibition of interferons. IL-1 mature GeneSeq EP456332Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, R14855lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Orencole &Dinarclio (1989) neutrophils and T lymphocytes, and/or Cytokine 1,14-20. inhibition of interferons. IL-1 beta GeneSeq WO9922763Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, Y08322lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Orencole &Dinarclio (1989) neutrophils and T lymphocytes, and/or Cytokine 1,14-20. inhibition of interferons. IL-3 variants GeneSeq WO8806161Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, P80382, P80383,lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancerP80384, and macrophages. Known functions include Interferens: APractical Approach, P80381 stimulating proliferation of immune cellsClemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Kitamura et al (1989) J Cell neutrophils and Tlymphocytes, and/or Physiol. 140 323-334. inhibition of interferons.IL-4 GeneSeq WO8702990 Interleukins are a group of Interleukin activitycan be determined inflammatory disorders, Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, P70615 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andSiegel & Mostowski (1990) J neutrophils and T lymphocytes, and/orImmunol Methods 132, 287-295. inhibition of interferons. IL-4 muteinsGeneSeq WO9747744 Interleukins are a group of Interleukin activity canbe determined inflammatory disorders, Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, W52151 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer W52152 macrophages. Known functions includeInterferens: A Practical Approach, W52153 stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, W52154 (e.g., T helpercells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225; W52155and lymphocytes), chemotaxis of and Siegel & Mostowski (1990) J W52156neutrophils and T lymphocytes, and/or Immunol Methods 132, 287-295.W52157 inhibition of interferons. W52158 W52159 W52160 W52161 W52162W52163 W52164 and W52165 IL-1 alpha GeneSeq EP324447 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,Accession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, P90108 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Orencole & Dinarello (1989) neutrophilsand T lymphocytes, and/or Cytokine 1, 14-20. inhibition of interferons.IL-3 variants GeneSeq WO9307171 Interleukins are a group of Interleukinactivity can be determined inflammatory disorders, Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, R38561, R38562, lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer R38563, R38564, macrophages.Known functions include Interferens: A Practical Approach, R38565,R38566, stimulating proliferation of immune cells Clemens et al.; eds,IRL Press, R38567, R38568, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; R38569, R38570, and lymphocytes),chemotaxis of and Aarden et al (1987) Eur. J. R38571, and neutrophilsand T lymphocytes, and/or Immunol 17, 1411-16. R38572 inhibition ofinterferons. IL-6 GeneSeq WO9402512 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, R45717 and lymphocytes, monocytes, and Matthewset al., in Lymphokines and cancer R45718 macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Aarden et al (1987) Eur. J. neutrophilsand T lymphocytes, and/or Immunol 17, 1411-16. inhibition ofinterferons. IL-13 GeneSeq WO9404680 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, R48624 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferens: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Boutelier et al (1995) J. neutrophils and Tlymphocytes, and/or Immunol. Methods 181, 29. inhibition of interferons.IL-4 mutein GeneSeq DE4137333 Interleukins are a group of Interleukinactivity can be determined inflammatory disorders, Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, R47182 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferens: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Siegel & Mostowski (1990) J neutrophils and Tlymphocytes, and/or Immunol Methods 132, 287-295. inhibition ofinterferons. IL-4 mutein GeneSeq DE4137333 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Y124XAccession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, R47183 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Siegel & Mostowski (1990) J neutrophilsand T lymphocytes, and/or Immunol Methods 132, 287-295. inhibition ofinterferons. IL-4 mutein GeneSeq DE4137333 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Y124GAccession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, R47184 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Siegel & Mostowski (1990) J neutrophilsand T lymphocytes, and/or Immunol Methods 132, 287-295. inhibition ofinterferons. Human GeneSeq WO9317698 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders,Interleukin-10 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, (precursor) R41664lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Thompson-Snipeset al (1991) J. neutrophils and T lymphocytes, and/or Exp. Med. 173,507-510. inhibition of interferons. Human GeneSeq WO9318783-AInterleukins are a group of Interleukin activity can be determinedinflammatory disorders, Interleukin-10 Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, R42642 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andThompson-Snipes et al (1991) J. neutrophils and T lymphocytes, and/orExp. Med. 173, 507-510. inhibition of interferons. Human GeneSeqEP569042 Interleukins are a group of interleukin activity can bedetermined inflammatory disorders, interleukin-1 Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, beta precursor. R42447 lymphocytes, monocytes,and Matthews et al., in Lymphokines and cancer macrophages. Knownfunctions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Orencole & Dinarello (1989)neutrophils and T lymphocytes, and/or Cytokine 1, 14-20. inhibition ofinterferons. Interleukin- GeneSeq EP578278 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, 1alphaAccession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, R45364 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225. andlymphocytes), chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. Human GeneSeq JP04063595 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,interleukin-3 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, variant R22814lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Kitamura et al(1989) J Cell neutrophils and T lymphocytes, and/or Physiol. 140323-334. inhibition of interferons. IL-1i fragments GeneSeq EP541920Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, R35484 andlymphocytes, monocytes, and Matthews et al., in Lymphokines and cancerR35485 macrophages. Known functions include Interferens: A PracticalApproach, stimulating proliferation of immune cells Clemens et al., eds,IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Orencole &Dinarclio (1989) neutrophils and T lymphocytes, and/or Cytokine 1,14-20. inhibition of interferons. IL-1 inhibitor GeneSeq EPS541920Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, (IL-li) Accession multifunctional cytokinessynthesized by using assays known in the art: immunologic disorders,R35486 and lymphocytes, monocytes, and Matthews et al., in Lymphokinesand cancer R35484 macrophages. Known functions include Interferens: APractical Approach, stimulating proliferation of immune cells Clemens etal., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andOrencole & Dinarclio (1989) neutrophils and T lymphocytes, and/orCytokine 1, 14-20. inhibition of interferons. ICE 22 kD subunit. GeneSeqEP533350 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, Accession multifunctional cytokinessynthesized by using assays known in the art: immunologic disorders,R33780 lymphocytes, monocytes, and Matthews et al., in Lymphokines andcancer macrophages. Known functions include Interferens: A PracticalApproach, stimulating proliferation of immune cells Clemens et al., eds,IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225. and lymphocytes), chemotaxis of neutrophils and Tlymphocytes, and/or inhibition of interferons. ICE 20 kD subunit.GeneSeq EP533350 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, Accession multifunctional cytokinessynthesized by using assays known in the art: immunologic disorders,R33781 lymphocytes, monocytes, and Matthews et al., in Lymphokines andcancer macrophages. Known functions include Interferens: A PracticalApproach, stimulating proliferation of immune cells Clemens et al., eds,IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225. and lymphocytes), chemotaxis of neutrophils and Tlymphocytes, and/or inhibition of interferons. ICE 10 kD GeneSeqEP533350 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, subunit Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, R33782 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225. and lymphocytes), chemotaxis ofneutrophils and T lymphocytes, and/or inhibition of interferons. HumanGeneSeq WO9317698 Interleukins are a group of Interleukin activity canbe determined inflammatory disorders, Interleukin-10 Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, (precursor) R41664 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Thompson-Snipes et al (1991) J.neutrophils and T lymphocytes, and/or Exp. Med. 173, 507-510. inhibitionof interferons. Human GeneSeq WO9318783 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders,Interleukin-10 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, R42642 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer macrophages.Known functions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Thompson-Snipes et al (1991) J.neutrophils and T lymphocytes, and/or Exp. Med. 173, 507-510. inhibitionof interferons. Human GeneSeq EP569042 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders,Interleukin-1 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, beta precursor R42447lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Kitamura et al(1989) J Cell neutrophils and T lymphocytes, and/or Physiol. 140323-334. inhibition of interferons. Human GeneSeq WO9403492 Interleukinsare a group of Interleukin activity can be determined inflammatorydisorders, interleukin-6 Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, R49041lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Aarden et al(1987) Eur. J. neutrophils and T lymphocytes, and/or Immunol 17,1411-16. inhibition of interferons. Mutant Interleukin 6 GeneSeqWO9411402 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, S176R Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, R54990 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andAarden et al (1987) Eur. J. neutrophils and T lymphocytes, and/orImmunol 17, 1411-16. inhibition of interferons. Interleukin 6 GeneSeqJP06145063 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, Accession multifunctional cytokinessynthesized by using assays known in the art: immunologic disorders,R55256 lymphocytes, monocytes, and Matthews et al., in Lymphokines andcancer macrophages. Known functions include Interferens: A PracticalApproach, stimulating proliferation of immune cells Clemens et al., eds,IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Aarden et al(1987) Eur. J. neutrophils and T lymphocytes, and/or Immunol 17,1411-16. inhibition of interferons. Interleukin 8 GeneSeq JP06100595Interleukins are a group of Interleukin activity can be determinedSoluble IL-8 receptor (IL-8) receptor Accession multifunctionalcytokines synthesized by using assays known in the art: polypeptides maybe R53932 lymphocytes, monocytes, and Matthews et al., in Lymphokinesand useful for inhibiting macrophages. Known functions includeInterferens: A Practical Approach, interleukin activities. stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Holmes et al (1991) Science 253,neutrophils and T lymphocytes, and/or 1278-80. inhibition ofinterferons. Human GeneSeq U.S. Pat. No. 5,328,988 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,interleukin-7 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, R59919 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer macrophages.Known functions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Park et al (1990) J. Exp. Med.neutrophils and T lymphocytes, and/or 171, 1073-79. inhibition ofinterferons. IL-3 containing GeneSeq WO9521254 Interleukins are a groupof Interleukin activity can be determined inflammatory disorders, fusionprotein. Accession multifunctional cytokines synthesized by using assaysknown in the art: immunologic disorders, R79342 and lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer R79344macrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Kitamura et al(1989) J Cell neutrophils and T lymphocytes, and/or Physiol. 140323-334. inhibition of interferons. IL-3 mutant GeneSeq ZA9402636Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, proteins Accession multifunctional cytokinessynthesized by using assays known in the art: immunologic disorders,R79254, R79255, lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer R79256, R79257, macrophages. Known functionsinclude Interferens: A Practical Approach, R79258, R79259, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, R79260,R79261, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; R79262, R79263, and lymphocytes), chemotaxis of andGiri et al (1994) EMBO J. 13 R79264, R79265, neutrophils and Tlymphocytes, and/or 2822-2830. R79266, R79267, inhibition ofinterferons. R79268, R79269, R79270, R79271, R79272, R79273, R79274,R79275, R79276, R79277, R79278, R79279, R79280, R79281, R79282, 879283,R79284, and R79285 IL-12 p40 GeneSeq AU9466072 Interleukins are a groupof Interleukin activity can be determined inflammatory disorders,subunit. Accession multifunctional cytokines synthesized by using assaysknown in the art: immunologic disorders, R63018 lymphocytes, monocytes,and Matthews et al., in Lymphokines and cancer macrophages. Knownfunctions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225.and lymphocytes), chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. AGF GeneSeq WO9429344 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,Accession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, R64240 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferens: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225. andlymphocytes), chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. Human GeneSeq WO9519786 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,interlaukin-12 40 kD Accession multifunctional cytokines synthesized byusing assays known in the art: immunologic disorders, subunit R79187lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Hori et al(1987), Blood 70, neutrophils and T lymphocytes, and/or 1069-1078.inhibition of interferons. Human GeneSeq WO9530695 Interleukins are agroup of Interleukin activity can be determined Soluble IL-8 receptorinterleukin-15 Accession multifunctional cytokines synthesized by usingassays known in the art: polypeptides may be receptor from R90843lymphocytes, monocytes, and Matthews et al., in Lymphokines and usefulfor inhibiting clone P1 macrophages. Known functions includeInterferens: A Practical Approach, interleukin activities. stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Giri et al (1994) EMBO J. 13neutrophils and T lymphocytes, and/or 2822-2830. inhibition ofinterferons. Human GeneSeq WO9604306 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders,interleukin-7 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, R92796 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer macrophages.Known functions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Park et al (1990) J. Exp. Med.neutrophils and T lymphocytes, and/or 171, 1073-79. inhibition ofinterferons. interleukin-9 GeneSeq WO9604306 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Accesionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, R92797 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferens: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Yang et al (1989) Blood 74, neutrophils and Tlymphocytes, and/or 1880-84. inhibition of interferons. interleukin-3GeneSeq WO9604306 Interleukins are a group of Interleukin activity canbe determined inflammatory disorders, Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, R92801 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andKitamura et al (1989) J Cell neutrophils and T lymphocytes, and/orPhysiol. 140 323-334. inhibition of interferons. Human GeneSeq WO9604306Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, interleukin-5 Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, R92802 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andKitamura et al (1989) J Cell neutrophils and T lymphocytes, and/orPhysiol. 140 323-334. inhibition of interferons. Recombinant GeneSeqDE19617202 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, interleukin-16 Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, W33373 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferens: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Lim et al (1996) J. Immunol. 156, neutrophils and Tlymphocytes, and/or 2566-70. inhibition of interferons. Human IL-16GeneSeq DE19617202 Interleukins are a group of Interleukin activity canbe determined inflammatory disorders, protein Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, W33234 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andLim et al (1996) J. Immunol. 156, neutrophils and T lymphocytes, and/or2566-70. inhibition of interferons. Thrl 17 human GeneSeq WO9708321Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, interleukin 9 Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, W27521 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferens:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225. and lymphocytes), chemotaxis ofneutrophils and T lymphocytes, and/or inhibition of interferons. Metl 17human GeneSeq WO9708321 Interleukins are a group of Interleukin activitycan be determined inflammatory disorders, interleukin 9 Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, W27522 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferens: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Yang et al (1989) Blood 74, neutrophils and Tlymphocytes, and/or 1880-84. inhibition of interferons. Human GeneSeqEP86-4585 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, intracellular IL-1 Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, receptor W77158 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer antagonist. macrophages.Known functions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Orencole & Dinarello (1989)neutrophils and T lymphocytes, and/or Cytokine 1, 14-20. inhibition ofinterferons. Human GeneSeq EP864585 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders,interleukin-18 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, protein (IL-18) W77158lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferens: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and USHIO et al(1996) J. Immunol. neutrophils and T lymphocytes, and/or 156, 4274-79.inhibition of interferons. Human GeneSeq EP861663 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,interleukin-18 Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, W77077 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer macrophages.Known functions include Interferens: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and USHIO et al (1996) J. Immunol.neutrophils and T lymphocytes, and/or 156, 4274-79. inhibition ofinterferons. Human interleukin GeneSeq EP861663 Interleukins are a groupof Interleukin activity can be determined inflammatory disorders, 18derivatives Accessions multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, W77083, lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer W77084,macrophages. Known functions include Interferons: A Practical Approach,W77085, stimulating proliferation of immune cells Clemens et al., eds,IRL Press, W77086, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; W77087, and lymphocytes), chemotaxisof and Ushio et al (1996) J. Immunol, W77088, and neutrophils and Tlymphocytes, and/or 156, 4274-79. W77089 inhibition of interferons.Interleukin-9 GeneSeq WO9827997 Interleukins are a group of Interleukinactivity can be determined inflammatory disorders, (IL-9) matureAccession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, protein (Thr117 W68158 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer version).macrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Yang et al (1989)Blood 74, neutrophils and T lymphocytes, and/or 1880-84. inhibition ofinterferons. IL-9 mature GenSeq Accession WO9827997 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,protein variant W68157 multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, (Met117 version)lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Yang et al (1989)Blood 74, neutrophils and T lymphocytes, and/or 1880-84. inhibition ofinterferons. Human IL-9 GeneSeq WO9824904 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, receptorprotein Accession multifunctional cytokines synthesized by using assaysknown in the art: immunologic disorders, variant #3. W64058 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer macrophages.Known functions include Interferons: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Yang et at (1989) Blood 74,neutrophils and T lymphocytes, and/or 1880-84. inhibition ofinterferons. Human IL-9 GenSeq Accession WO9824904 Interleukins are agroup of Interleukin activity can be determined Soluble IL-9 receptorreceptor protein W64060 multifunctional cytokines synthesized by usingassays known in the art: polypeptides may be variant fragmentlymphocytes, monocytes, and Matthews et al., in Lymphokines and usefulfor inhibiting macrophages. Known functions include Interferons: APractical Approach, interleukin activities. stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Yang et al (1989) Blood 74, neutrophilsand T lymphocytes, and/or 1880-84. inhibition of interferons. Human IL-9GeneSeq WO9824904 Interleukins are a group of Interleukin activity canbe determined Soluble IL-9 receptor receptor protein Accessionmultifunctional cytokines synthesized by using assays known in the art:polypeptides may be variant #3. W64061 lymphocytes, monocytes, andMatthews et al., in Lymphokines and useful for inhibiting macrophages.Known functions include Interferons: A Practical Approach, interleukinactivities. stimulating proliferation of immune cells Clemens et al.,eds, IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington,D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of and Yang et at(1989) Blood 74, neutrophils and T lymphocytes, and/or 1880-84.inhibition of interferons. Human GeneSeq WO9817689 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,Interleukin-12 p40 Accession multifunctional cytokines synthesized byusing assays known in the art: immunologic disorders, protein W51311lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Hori et al(1987), Blood 70, neutrophils and T lymphocytes, and/or 1069-1078.inhibition of interferons. Human GeneSeq WO9817689 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,Interleukin-12 p35 Accession multifunctional cytokines synthesized byusing assays known in the art: immunologic disorders, protein W51312lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Hori et al(1987), Blood 70, neutrophils and T lymphocytes, and/or 1069-1078.inhibition of interferons. Human protein GeneSeq DE19649233 Interleukinsare a group of Interleukin activity can be determined inflammatorydisorders, with IL-16 activity Accession multifunctional cytokinessynthesized by using assays known in the art: immunologic disorders,W63753 lymphocytes, monocytes, and Matthews et al., in Lymphokines andcancer macrophages. Known functions include Interferons: A PracticalApproach, stimulating proliferation of immune cells Clemens et al., eds,IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Lim et al (1996)J. Immunol. 156, neutrophils and T lymphocytes, and/or 2566-70.inhibition of interferons. Human protein GeneSeq DE19649233-Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, with IL-16 activity Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, W59425 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferons:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andLim et al (1996) J. Immunol. 156, neutrophils and T lymphocytes, and/or2566-70. inhibition of interferons. Human GeneSeq U.S. Pat. No.5,747,024 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, interleukin-15 Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, W53878 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferons: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Giri et al (1994) EMBO J. 13 neutrophils and Tlymphocytes, and/or 2822-2830. inhibition of interferons. Humanwild-type GeneSeq WO9747744 Interleukins are a group of Interleukinactivity can be determined inflammatory disorders, interleukin-4 (hIL-Accession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, 4) protein W52149 lymphocytes,monocytes, and Matthews et al., in Lymphokines and cancer macrophages.Known functions include Interferons: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Siegel & Mostowski (1990) Jneutrophils and T lymphocytes, and/or Immunol Methods 132, 287-295.inhibition of interferons. interleukin-4 GeneSeq WO9747744 Interleukinsare a group of Interleukin activity can be determined inflammatorydisorders, muteins Accessions multifunctional cytokines synthesized byusing assays known in the art: immunologic disorders, W52150,lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancerW52151, macrophages. Known functions include Interferons: A PracticalApproach, W52153, stimulating proliferation of immune cells Clemens etal., eds, IRL Press, W52154, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; W52155, andlymphocytes), chemotaxis of and Siegel & Mostowski (1990) J W52156,neutrophils and T lymphocytes, and/or Immunol Methods 132, 287-295.W52157, inhibition of interferons. W52158, W52159, W52160, W52161,W52162, W52163, W52164, W52165, W52166, and W52167 Human interleukinGeneSeq WO9935268 Interleukins are a group of Interleukin activity canbe determined inflammatory disorders, 1 delta Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, Y28408 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferons:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andOrencole & Dinarello (1989) neutrophils and T lymphocytes, and/orCytokine 1, 14-20. inhibition of interferons. Human GeneSeq WO9935268Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, interleukin-1 Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, receptor antagonist Y24395 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer beta macrophages. Knownfunctions include Interferons: A Practical Approach, stimulatingproliferation of immune cells Clemens et al., eds, IRL, Press, (e.g., Thelper cells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225;and lymphocytes), chemotaxis of and Orencole & Dinarello (1989)neutrophils and T lymphocytes, and/or Cytokine 1, 14-20. inhibition ofinterferons. Human EDIRF II GeneSeq WO9932632 Interleukins are a groupof Interleukin activity can be determined inflammatory disorders,protein sequence Accession multifunctional cytokines synthesized byusing assays known in the art: immunologic disorders, Y22199lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225. and lymphocytes), chemotaxis of neutrophils and Tlymphocytes, and/or inhibition of interferons. Human EDIRF I GeneSeqWO9932632 Interleukins are a group of Interleukin activity can bedetermined inflammatory disorders, protein sequence Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, Y22197 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferons: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cell, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225. and lymphocytes),chemotaxis of neutrophils and T lymphocytes, and/or inhibition ofinterferons. Human IL-1RD10 GeneSeq WO9919480 Interleukins are a groupof Interleukin activity can be determined Soluble IL-1RD10 proteinsequence Accession multifunctional cytokines synthesized by using assaysknown in the art: receptor polypeptides Y14131 lymphocytes, monocytes,and Matthews et al., in Lymphokines and may be useful for macrophages.Known functions include Interferons: A Practical Approach, inhibitinginterleukin stimulating proliferation of immune cells Clemens et al.,eds, IRL Press, activites. (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andOrencole & Dinarello (1989) neutrophils and T lymphocytes, and/orCytokine 1, 14-20. inhibition of interferons. Human IL-1RD9 GeneSeqWO9919480 Interleukins are a group of Interleukin activity can bedetermined Soluble IL-1RD10 Accession multifunctional cytokinessynthesized by using assays known in the art: receptor polypeptidesY14122 lymphocytes, monocytes, and Matthews et al., in Lymphokines andmay be useful for macrophages. Known functions include Interferons: APractical Approach, inhibiting interleukin stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, activites. (e.g., T helpercells, B cells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Orencole & Dinarello (1989) neutrophilsand T lymphocytes, and/or Cytokine 1, 14-20. inhibition of interferons.Human DNAX GeneSeq WO9919491 Interleukins are a group of Interleukinactivity can be determined inflammatory disorders, interleukin-40Accession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, Y09196 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferons: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225. andlymphocytes), chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. (DIL-40) GeneSeq WO9919491 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,alternative Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, sequence Y09197lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225. and lymphocytes), chemotaxis of neutrophils and Tlymphocytes, and/or inhibition of interferons. IL-11 GeneSeq WO9405318Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, R50176lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Lu et al (1994) Jimmunol. neutrophils and T lymphocytes, and/or Methods 173, 19.inhibition of interferons. Human GeneSeq EP566410 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,adipogenesis Accession multifunctional cytokines synthesized by usingassays known in the art: immunologic disorders, inhibitory factor R43260lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225. and lymphocytes), chemotaxis of neutrophils and Tlymphocytes, and/or inhibition of interferons. IL-11 GeneSeq JP08127539Interleukins are a group of Interleukin activity can be determinedinflammatory disorders, Accession multifunctional cytokines synthesizedby using assays known in the art: immunologic disorders, W02202lymphocytes, monocytes, and Matthews et al., in Lymphokines and cancermacrophages. Known functions include Interferons: A Practical Approach,stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Lu et al (1994) Jimmunol. neutrophils and T lymphocytes, and/or Methods 173, 19.inhibition of interferons. IL-14 GeneSeq WO9416074 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,Accession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, R55800 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferons: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Ambrus et al (1993) PNAS 90, neutrophilsand T lymphocytes, and/or 63330-34. inhibition of interferons. IL-17receptor GeneSeq U.S. Pat. No. 6,072,033 Interleukins are a group ofInterleukin activity can be determined Soluble IL-17 receptor Accessionmultifunctional cytokines synthesized by using assays known in the art:polypeptides may be B03807 lymphocytes, monocytes, and Matthews et al.,in Lymphokines and useful for inhibiting macrophages. Known functionsinclude Interferons: A Practical Approach, interleukin activities.stimulating proliferation of immune cells Clemens et al., eds, IRLPress, (e.g., T helper cells, B cells, eosinophils, Washington, D.C.1987, pp. 221-225; and lymphocytes), chemotaxis of and Yao et al (1995)J. Immunol. 155, neutrophils and T lymphocytes, and/or 5483-86.inhibition of interferons. IL-17 GeneSeq WO9518826 Interleukins are agroup of Interleukin activity can be determined inflammatory disorders,Accession multifunctional cytokines synthesized by using assays known inthe art: immunologic disorders, R76573 lymphocytes, monocytes, andMatthews et al., in Lymphokines and cancer macrophages. Known functionsinclude Interferons: A Practical Approach, stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Yao et al (1995) J. Immunol. 155,neutrophils and T lymphocytes, and/or 5483-86. inhibition ofinterferons. CTLA-8 GeneSeq WO9704097 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, W13651 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferons: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225. and lymphocytes),chemotaxis of neutrophils and T lymphocytes, and/or inhibition ofinterferons. IL-19 GeneSeq WO9808870 Interleukins are a group ofInterleukin activity can be determined inflammatory disorders, Accessionmultifunctional cytokines synthesized by using assays known in the art:immunologic disorders, W37935 lymphocytes, monocytes, and Matthews etal., in Lymphokines and cancer macrophages. Known functions includeInterferons: A Practical Approach, stimulating proliferation of immunecells Clemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Gallagher et al (2000) Genes neutrophils and Tlymphocytes, and/or Immun. 1, 442-50. inhibition of interferons. IL-21(TIF) GeneSeq WO0024758 Interleukins are a group of Interleukin activitycan be determined inflammatory disorders, Accession multifunctionalcytokines synthesized by using assays known in the art: immunologicdisorders, Y92879 lymphocytes, monocytes, and Matthews et al., inLymphokines and cancer macrophages. Known functions include Interferons:A Practical Approach, stimulating proliferation of immune cells Clemenset al., eds, IRL Press, (e.g., T helper cells, B cells, eosinophils,Washington, D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of andParrish-Novak et al (2000) neutrophils and T lymphocytes, and/or Nature408, 57-63. inhibition of interferons. IL-8 receptor GeneSeq WO9306229Interleukins are a group of Interleukin activity can be determinedSoluble IL-8 receptor Accession multifunctional cytokines synthesized byusing assays known in the art: polypeptides may be R33420 lymphocytes,monocytes, and Matthews et al., in Lymphokines and useful for inhibitingmacrophages. Known functions include Interferons: A Practical Approach,interleukin activities. stimulating proliferation of immune cellsClemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Holmes et al (1991) Science 253, neutrophils and Tlymphocytes, and/or 1278-80.. inhibition of interferons. Human type IIGeneSeq U.S. Pat. No. 5,464,937 Interleukins are a group of Interleukinactivity can be determined Soluble type II interleukin-1 Accessionmultifunctional cytokines synthesized by using assays known in the art:interleukin-1 receptor receptor R85480 lymphocytes, monocytes, andMatthews et al., in Lymphokines and polypeptides may be macrophages.Known functions include Interferons: A Practical Approach, useful forinhibiting stimulating proliferation of immune cells Clemens et al.,eds. IRL Press, interleukin activities. (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Orencole & Dinarello (1989) neutrophils and Tlymphocytes, and/or Cytokine 1, 14-20. inhibition of interferons. HumanGeneSeq EP638644 Interleukins are a group of Interleukin activity can bedetermined Soluble IL-12 receptor interleukin-12 Accessionmultifunctional cytokines synthesized by using assays known in the art:polypeptides may be receptor R69632 lymphocytes, monocytes, and Matthewset al., in Lymphokines and useful for inhibiting macrophages. Knownfunctions include Interferons: A Practical Approach, interleukinactivities. stimulating proliferation of immune cells Clemens et al.,eds, IRL Press, (e.g., T helper cells, B cells, eosinophils, Washington,D.C. 1987, pp. 221-225; and lymphocytes), chemotaxis of and Hori et al(1987), Blood 70, neutrophils and T lymphocytes, and/or 1069-1078.inhibition of interferons. Interleukin 8 GeneSeq U.S. Pat. No. 5,440,021Interleukins are a group of Interleukin activity can be determinedSoluble IL-8 receptor B receptor B Accession multifunctional cytokinessynthesized by using assays known in the art: polypeptides may be R80758lymphocytes, monocytes, and Matthews et al., in Lymphokines and usefulfor inhibiting macrophages. Known functions include Interferons: APractical Approach, interleukin activities. stimulating proliferation ofimmune cells Clemens et al., eds, IRL Press, (e.g., T helper cells, Bcells, eosinophils, Washington, D.C. 1987, pp. 221-225; andlymphocytes), chemotaxis of and Holmes et al (1991) Science 253,neutrophils and T lymphocytes, and/or 1278-80. inhibition ofinterferons. Human IL-8 GeneSeq JP08103276 Interleukins are a group ofInterleukin activity can be determined Soluble IL-8 receptor A receptorprotein Accession B09989 multifunctional cytokines synthesized by usingassays known in the art: polypeptides may be hIL8RA lymphocytes,monocytes, and Matthews et al., in Lymphokines and useful for inhibitingmacrophages. Known functions include Interferons: A Practical Approach,interleukin activities. stimulating proliferation of immune cellsClemens et al., eds, IRL Press, (e.g., T helper cells, B cells,eosinophils, Washington, D.C. 1987, pp. 221-225; and lymphocytes),chemotaxis of and Holmes et al (1991) Science 253, neutrophils and Tlymphocytes, and/or 1278-80. inhibition of interferons. Human IL-8GeneSeq JP08103276 Interleukins are a group of Interleukin activity canbe Soluble IL-8 receptor receptor protein Accession multifunctionalcytokines synthesized determined using asays known in the polypeptidesmay be hIL8R B09990 by lymphocytes, monocytes, and art: Matthews et al.,in Lymphokines useful for inhibiting macrophages. Known functionsinclude and Interferons: A Practical interleukin activities. stimulatingproliferation of immune Approach, Clemens et al., eds, IRL cells (e.g.,T helper cells, B cells, Press, Washington, D.C. 1987, pp. eosinophils,and lymphocytes), 221-225; and Holmes et al (1991) chemotaxis ofneutrophils and T Science 253, 1278-80. lymphocytes, and/or inhibitionof interferons. Interleukin-2 GeneSeq WO9621732 Interleukins are a groupof Interleukin activity can be Soluble IL-2 receptor receptor associatedAccession multifunctional cytokines synthesized determined using assaysknown in polypeptides may be protein p43 R97569 by lymphocytes,monocytes, and the art: Matthews et al., in useful for inhibitingmacrophages. Known functions include Lymphokines and Interferons: Ainterleukin activities. stimulating proliferation of immune PracticalApproach, Clemens et al., cells (e.g., T helper cells, B cells, eds, IRLPress, Washington, D.C. eosinophils, and lymphocytes), 1987, pp.221-225; and Gillis et al chemotaxis of neutrophils and T (1978) J.Immunol. 120, 2027. lymphocytes, and/or inhibition of interferons. HumanGeneSeq WO9629408 Interleukins are a group of Interleukin activity canbe Soluble IL-17 receptor interleukin-17 Accession multifunctionalcytokines synthesized determined using assays known in polypeptides maybe receptor W04185 by lymphocytes, monocytes, and the art: Matthews etal., in useful for inhibiting macrophages. Known functions includeLymphokines and Interferons: A interleukin activities. stimulatingproliferation of immune Practical Approach, Clemens et al., cells (e.g.,T helper cells, B cells, eds, IRL Press, Washington, D.C. eosinophils,and lymphocytes), 1987, pp. 221-225; and Yao et al chemotaxis ofneutrophils and T (1995) J. Immunol. 155, 5483-86. lymphocytes, and/orinhibition of interferons. Human GeneSeq WO9619574 Interleukins are agroup of Interleukin activity can be Soluble IL-11 receptorinterleukin-11 Accession multifunctional cytokines synthesizeddetermined using assays known in polypeptides may be receptor R99090 bylymphocytes, monocytes, and the art: Matthews et al., in useful forinhibiting macrophages. Known functions include Lymphokines andInterferons: A interleukin activities. stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Lu et al chemotaxis of neutrophils and T (1994) Jimmunol. Methods 173, 19. lymphocytes, and/or inhibition of interferons.Human GeneSeq WO9623067 Interleukins are a group of Interleukin activitycan be Inflammatory disorders, interleukin-1 Accession multifunctionalcytokines synthesized determined using assays known in immunologicdisorders, receptor accessory W01911 by lymphocytes, monocytes, and theart: Matthews et al., in cancer protein macrophages. Known functionsinclude Lymphokines and Interferons: A stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Orencole & chemotaxis of neutrophils and TDinarello (1989) Cytokine 1, 14-20. lymphocytes, and/or inhibition ofinterferons. AGF Protein GeneSeq U.S. Pat. No. 5,488,032 Interleukinsare a group of Interleukin activity can be Inflammatory disorders,Accession multifunctional cytokines synthesized determined using assaysknown in immunologic disorders, R92749 by lymphocytes, monocytes, andthe art: Matthews et al., in cancer macrophages. Known functions includeLymphokines and Interferons: A stimulating proliferation of immunePractical Approach, Clemens et al., cells (e.g., T helper cells, Bcells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225. chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. Human GeneSeq W09607739 Interleukins are agroup of Interleukin activity can be Soluble IL-type-3 interleukin-1type- Accession multifunctional cytokines synthesized determined usingassays known in receptor polypeptides 3 receptor R91064 by lymphocytes,monocytes, and the art: Matthews et al., in may be useful formacrophages. Known functions include Lymphokines and Interferons: Ainhibiting interleukin stimulating proliferation of immune PracticalApproach, Clemens et al., activities cells (e.g., T helper cells, Bcells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Orencole & chemotaxis of neutrophils and TDinarello (1989) Cytokine 1, 14-20. lymphocytes, and/or inhibition ofinterferons. Human GeneSeq WO9720926 Interleukins are a group ofInterleukin activity can be Soluble IL-13 beta interleukin-13 betaAccession multifunctional cytokines synthesized determined using assaysknown in receptor polypeptides receptor W24972 by lymphocytes,monocytes, and the art: Matthews et al., in may be useful formacrophages. Known functions include Lymphokines and Interferons: Ainhibiting interleukin stimulating proliferation of immune PracticalApproach, Clemens et al., activities. cells (e.g., T helper cells, Bcells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Boutelier et chemotaxis of neutrophils and T al(1995) J. Immunol. Methods lymphocytes, and/or inhibition of 181, 29.interferons. Human GeneSeq WO9720926 Interleukins are a group ofInterleukin activity can be Soluble IL-13 alpha interleukin-13 Accessionmultifunctional cytokines synthesized determined using assays known inreceptor polypeptides alpha receptor W24973 by lymphocytes, monocytes,and the art: Matthews et al., in may be useful for macrophages. Knownfunctions include Lymphokines and Interferons: A inhibiting interleukinstimulating proliferation of immune Practical Approach, Clemens et al.,activities. cells (e.g., T helper cells, B cells, eds, IRL Press,Washington, D.C. eosinophils, and lymphocytes), 1987, pp. 221-225; andBoutelier et chemotaxis of neutrophils and T al (1995) J. Immunol.Methods lymphocytes, and/or inhibition of 181, 29. interferons. HumanGeneSeq U.S. Pat. No. 5,599,905 Interleukins are a group of Interleukinactivity can be Soluble IL-4 receptor interleukin-4 Accessionmultifunctional cytokines synthesized determined using assays known inpolypeptides may be receptor W13499 by lymphocytes, monocytes, and theart: Matthews et al., in useful for inhibiting macrophages. Knownfunctions include Lymphokines and Interferons: A interleukin activities.stimulating proliferation of immune Practical Approach, Clemens et al.,cells (e.g., T helper cells, B cells, eds, IRL Press, Washington, D.C.eosinophils, and lymphocytes), 1987, pp. 221-225; and Siegel &chemotaxis of neutrophils and T Mostowski (1990) J Immunol lymphocytes,and/or inhibition of Methods 132, 287-295. interferons. Human GeneSeqEP759466 Interleukins are a group of Interleukin activity can be SolubleIL-12 beta-2 interleukin-12 Accession multifunctional cytokinessynthesized determined using assays known in receptor polypeptidesbeta-2 receptor W12771 by lymphocytes, monocytes, and the art: Matthewset al., in may be useful for macrophages. Known functions includeLymphokines and Interferons: A inhibiting interleukin stimulatingproliferation of immune Practical Approach, Clemens et al., activities.cells (e.g., T helper cells, B cells, eds, IRL Press, Washington, D.C.eosinophils, and lymphocytes), 1987, pp. 221-225; and Hori et alchemotaxis of neutrophils and T (1987), Blood 70, 1069-1078.lymphocytes, and/or inhibition of interferons. Human GeneSeq EP759466Interleukins are a group of Interleukin activity can be Soluble IL-12beta-1 interleukin-12 Accession multifunctional cytokines synthesizeddetermined using assays known in receptor polypeptides beta-1 receptor.W12772 by lymphocytes, monocytes, and the art: Matthews et al., in maybe useful for macrophages. Known functions include Lymphokines andInterferons: A inhibiting interleukin stimulating proliferation ofimmune Practical Approach, Clemens et al., activities. cells (e.g., Thelper cells, B cells, eds, IRL Press, Washington, D.C. eosinophils, andlymphocytes), 1987, pp. 221-225; and Hori et at chemotaxis ofneutrophils and T (1987), Blood 70, 1069-1078. lymphocytes, and/orinhibition of interferons. Human IL-9 GeneSeq WO9824904 Interleukins area group of Interleukin activity can be Soluble IL-9 receptor receptorprotein Accessions multifunctional cytokines synthesized determinedusing assays known in polypeptides may be W64055, by lymphocytes,monocytes, and the art: Matthews et al., in useful for inhibitingW64056, and macrophages. Known functions include Lymphokines andInterferons: A interleukin activities. W64057 stimulating proliferationof immune Practical Approach, Clemens et al., cells (e.g., T helpercells, B cells, eds, IRL Press, Washington, D.C. eosinophils, andlymphocytes), 1987, pp. 221-225; and Yang et al chemotaxis ofneutrophils and T (1989), Blood 74, 1880-84.. lymphocytes, and/orinhibition of interferons. IL-10 receptor GeneSeq U.S. Pat. No.5,716,804 Interleukins are a group of Interleukin activity can beSoluble IL-10 receptor Accession multifunctional cytokines synthesizeddetermined using assays known in polypeptides may be W41804 bylymphocytes, monocytes, and the art: Matthews et al., in useful forinhibiting macrophages. Known functions include Lymphokines andInterferons: A interleukin activities. stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Thompson- chemotaxis of neutrophils and T Snipeset al (1991) J. Exp. Med. 173, lymphocytes, and/or inhibition of507-510. interferons. Human IL-6 GeneSeq JP11196867 Interleukins are agroup of Interleukin activity can be Soluble IL-6 receptor receptorAccession multifunctional cytokines synthesized determined using assaysknown in polypeptides may be Y30938 by lymphocytes, monocytes, and theart: Matthews et al., in useful for inhibiting macrophages. Knownfunctions include Lymphokines and Interferons: A interleukin activities.stimulating proliferation of immune Practical Approach, Clemens et al.,cells (e.g., T helper cells, B cells, eds, IRL Press, Washington, D.C.eosinophils, and lymphocytes), 1987, pp. 221-225; and Aarden et alchemotaxis of neutrophils and T (1987) Eur. J. Immunol 17, 1411-16.lymphocytes, and/or inhibition of interferons. Il-17 receptor GeneSeqU.S. Pat. No. 6,096,305 Interleukins are a group of Interleukin activitycan be Soluble IL-17 receptor Accession multifunctional cytokinessynthesized determined using assays known in polypeptides may be Y97181by lymphocytes, monocytes, and the art: Matthews et al., in useful forinhibiting macrophages. Known functions include Lymphokines andInterferons: A interleukin activities. stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Yao et al chemotaxis of neutrophils and T (1995)J. Immunol. 155, 5483-86. lymphocytes, and/or inhibition of interferons.Il-17 receptor GeneSeq U.S. Pat. No. 6,100,235 Interleukins are a groupof Interleukin activity can be Soluble IL-17 receptor Accessionmultifunctional cytokines synthesized determined using assays known inpolypeptides may be Y97131 by lymphocytes, monocytes, and the art:Matthews et al., in useful for inhibiting macrophages. Known functionsinclude Lymphokines and Interferons: A interleukin activities.stimulating proliferation of immune Practical Approach, Clemens et al.,cells (e.g., T helper cells, B cells, eds, IRL Press, Washington, D.C.eosinophils, and lymphocytes), 1987, pp. 221-225; and Yao et alchemotaxis of neutrophils and T (1995) J. Immunol. 155, 5483-86.lymphocytes, and/or inhibition of interferons. Human GeneSeq EP509826Interleukins are a group of Interleukin activity can be Soluble IL-3receptor interleukin-3 Accession multifunctional cytokines synthesizeddetermined using assays known in polypeptides may be receptor R25300 bylymphocytes, monocytes, and the art: Matthews et al., in useful forinhibiting macrophages. Known functions include Lymphokines andInterferons: A interleukin activities. stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL, Press, Washington, D.C. eosinophils, andlymphocytes), 1987, pp. 221-225; and Kitamura et chemotaxis ofneutrophils and T al (1989) J Cell Physiol. 140 323-334. lymphocytes,and/or inhibition of interferons. Human GM-CSF GeneSeq WO9102063Interleukins are a group of Interleukin activity can be Soluble GM-CSFreceptor Accession multifunctional cytokines synthesized determinedusing assays known in receptor polypeptides R10919 by lymphocytes,monocytes, and the art: Matthews et al., in may be useful formacrophages. Known functions include Lymphokines and Interferons: Ainhibiting interleukin stimulating proliferation of immune PracticalApproach, Clemens et al., activities. cells (e.g., T helper cells, Bcells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225. chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. Human IL-5 GeneSeq EP492214 Interleukins area group of Interleukin activity can be Soluble IL-5 receptor receptoralpha Accession multifunctional cytokines synthesized determined usingassays known in alpha polypeptides may chain R25064 by lymphocytes,monocytes, and the art: Matthews et al., in be useful for inhibitingmacrophages. Known functions include Lymphokines and Interferons: Ainterleukin activities. stimulating proliferation of immune PracticalApproach, Clemens et al., cells (e.g., T helper cells, B cells, eds, IRLPress, Washington, D.C. eosinophils, and lymphocytes), 1987, pp.221-225; and Kitamura et chemotaxis of neutrophils and T al (1989) JCell Physiol. 140, 323-334. lymphocytes, and/or inhibition ofinterferons. Il-5 receptor GeneSeq WO9847923 Interleukins are a group ofInterleukin activity can be Soluble IL-5 receptor Accessionmultifunctional cytokines synthesized determined using assays known inpolypeptides may be W82842 by lymphocytes, monocytes, and the art:Matthews et al., in useful for inhibiting macrophages. Known functionsinclude Lymphokines and Interferons: A interleukin activities.stimulating proliferation of immune Practical Approach, Clemens et al.,cells (e.g., T helper cells, B cells, eds, IRL Press, Washington, D.C.eosinophils, and lymphocytes), 1987, pp. 221-225; and Kitamura etchemotaxis of neutrophils and T al (1989) J Cell Physiol. 140, 323-334.lymphocytes, and/or inhibition of interferons. Il-6 receptor GeneSeqJP05091892 Interleukins are a group of Interleukin activity can beSoluble IL-6 receptor Accession multifunctional cytokines synthesizeddetermined using assays known in polypeptides may be R37215 bylymphocytes, monocytes, and the art: Matthews et al., in useful forinhibiting macrophages. Known functions include Lymphokines andInterferons: A interleukin activities. stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Aarden et al chemotaxis of neutrophils and T(1987) Eur. J. Immunol 17, 1411-16. lymphocytes, and/or inhibition ofinterferons. Human B cell GeneSeq AU8928720 Interleukins are a group ofInterleukin activity can be Soluble B cell stimulating factor- Accessionmultifunctional cytokines synthesized determined using assays known instimulating factor-2 2 receptor P90525 by lymphocytes, monocytes, andthe art: Matthews et al., in receptor polypeptides macrophages. Knownfunctions include Lymphokines and Interferons: A may be useful forstimulating proliferation of immune Practical Approach, Clemens et al.,inhibiting interleukin cells (e.g., T helper cells, B cells, eds, IRLPress, Washington, D.C. activities. eosinophils, and lymphocytes), 1987,pp. 221-225. chemotaxis of neutrophils and T lymphocytes, and/orinhibition of interferons. IL-7 receptor clone GeneSeq EP403114Interleukins are a group of Interleukin activity can be Soluble IL-7receptor Accession multifunctional cytokines synthesized determinedusing assays known in polypeptides may be R08330 by lymphocytes,monocytes, and the art: Matthews et al., in useful for inhibitingmacrophages. Known functions include Lymphokines and Interferons: Ainterleukin activities. stimulating proliferation of immune PracticalApproach, Clemens et al., cells (e.g., T helper cells, B cells, eds, IRLPress, Washington, D.C. eosinophils, and lymphocytes), 1987, pp.221-225; and Park et al chemotaxis of neutrophils and T (1990) J. Exp.Med. 171, 1073-79. lymphocytes, and/or inhibition of interferons. EPOreceptor; GeneSeq WO9008822 EPO Receptor is involved in the EPO Receptoractivity can be inflammatory disorders, EPOR Accession proliferation anddifferentiation of determined using assays known in immunologicdisorders, R06512 erythroblasts. the art, such as, J Biol Chem 2001cancer, erythroblast Mar 23; 276(12: 8995-9002; JAK2 proliferation andprotein tyrosine kinase activity: differentiation Blood 1994 Sep 1;84(5): 1501-7 and Mol Cell Biol. 1994 Oct; 14(10: 6506-14. IL-15receptor GeneSeq WO9530695 Interleukins are a group of Interleukinactivity can be Soluble IL-15 receptor Accession multifunctionalcytokines synthesized determined using assays known in polypeptides maybe R90843 by lymphocytes, monocytes, and the art: Matthews et al., inuseful for inhibiting macrophages. Known functions include Lymphokinesand Interferons: A interleukin activities. stimulating proliferation ofimmune Practical Approach, Clemens et al., cells (e.g., T helper cells,B cells, eds, IRL Press, Washington, D.C. eosinophils, and lymphocytes),1987, pp. 221-225; and Giri et al chemotaxis of neutrophils and T (1994)EMBO J. 13 2822-2830. lymphocytes, and/or inhibition of interferons.CD137; 4-1BB GeneSeq WO9507984 Activities associated with apoptosis,Apoptosis activity, NF-kB Soluble 4-1BB receptor Receptor ProteinAccession NF-kB activation, and co-stimulation of activation, and B andT cell co- polypeptides may be R70977 immune cells such as T and Bcells. stimulation can be determined using useful for inhibiting assaysknown in the art: Moore et apoptosis, NF-kB al., 1999, Science,285(5425): 260-3; activation, and/or co- Song HY et al., 1997 Proc NatlAcad stimulation of immune Sci USA 94(18): 9792-6; Epsevik cells such asB and T and Nissen-Meyer, 1986, J. cells. Immunol. Methods. BCMA GeneSeqWO0068378 Activities associated with apoptosis, Apoptosis activity,NF-kB Soluble BCMA receptor Accession NF-kB activation, andco-stimulation of activation, and B and T cell co- polypeptides may beY71979 immune cells such as T and B cells. stimulation can be determinedusing useful for inhibiting assays known in the art: Moore et apoptosis,NF-kB al., 1999, Science, 285(5425): 260-3; activation, and/or co- SongHY et al., 1997 Proc Natl Acad stimulation of immune Sci USA 94(18):9792-6; Epsevik cells such as B and T and Nissen-Meyer, 1986, J. cells.Immunol. Methods. CD27 GeneSeq WO9201049 Activities associated withapoptosis, Apoptosis activity, NF-kB Soluble CD27 Accession NF-kBactivation, and co-stimulation of activation, and B and T cell co-polypeptides may be R20814 immune cells such as T and B cells.stimulation can be determined using useful for inhibiting assays knownin the art: Moore et apoptosis, NF-kB al., 1999, Science, 285(5425):260-3; activation, and/or co- Song HY et al., 1997 Proc Natl Acadstimulation of immune Sci USA 94(18): 9792-6; Epsevik cells such as Band T and Nissen-Meyer, 1986, J. cells. Immunol. Methods. CD30 GeneSeqDE4200043 Activities associated with apoptosis, Apoptosis activity,NF-kB Soluble CD30 Accession NF-kB activation, and co-stimulation ofactivation, and B and T cell co- polypeptides may be R35478 immune cellssuch as T and B cells. stimulation can be determined using useful forinhibiting assays known in the art: Moore et apoptosis, NF-kB al., 1999,Science, 285(5425): 260-3; activation, and/or co- Song HY et al., 1997Proc Natl Acad stimulation of immune Sci USA 94(18): 9792-6; Epsevikcells such as B and T and Nissen-Meyer, 1986, J. cells. Immunol.Methods. CD40 GeneSeq WO9945944 Activities associated with apoptosis,Apoptosis activity, NF-kB Soluble CD40 Accession NF-kB activation, andco-stimulation of activation, and B and T cell polypeptides may beY33499 immune cells such as T and B cells. co-stimulation can bedetermined useful for inhibiting using assays known in the art:apoptosis, NF-kB Moore et al., 1999, Science activation, and/or co-285(5425): 260-3; Song HY et al., stimulation of immune 1997 Proc NatlAcad Sci USA cells such as B and T 94(18): 9792-6; Epsevik and cells.Nissen-Meyer, 1986, J. Immunol. Methods. EDAR Genbank Activitiesassociated with apoptosis, Apoptosis activity, NF-kB activation, ImmuneDisorders, Accession NF-kB activation, and co-stimulation of and B and Tcell co-stimulation can Lymphomas, X-linked AAD50077 immune cells suchas T and B cells. be determined using assays known in hypohidroticectodermal the art: Moore et al., 1999, Science, dysplasia 285(5425):260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18): 9792-6;Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. OX40; ACT-4GeneSeq WO9512673 Activities associated with apoptosis, Apoptosisactivity, NF-kB activation, Immune Disorders, Accession R74737 NF-kBactivation, and co-stimulation of and B and T cell co-stimulation canLymphomas, T cell immune cells such as T and B cells. be determinedusing assays known in disorders the art: Moore et al., 1999, Science,285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18):9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. TAClGeneSeq WO9839361 Activities associated with apoptosis, Apoptosisactivity, NF-kB activation, Soluble TACl receptor Accession NF-kBactivation, and co-stimulation of and B and T cell co-stimulation canpolypeptides may be W75783 immune cells such as T and B cells. bedetermined using assays known in useful for inhibiting the art: Moore etal., 1999, Science, apoptosis, NF-kB 285(5425): 260-3; Song HY et al.,activation, and/or co- 1997 Proc Natl Acad Sci USA stimulation of immune94(18): 9792-6; Epsevik and Nissen- cells such as B and T Meyer, 1986,J. Immunol. Methods. cells. TNF-R GeneSeq AU9058976 Activitiesassociates with apoptosis, Apoptosis activity, NF-kB Soluble TNF-Rreceptor Accession R10986 NF-kB activation, and co-stimulation ofactivation, and B and T cell co- polypeptides may be immune cells suchas T and B cells. stimulation can be determined using useful forinhibiting assays known in the art: Moore et al., apoptosis, NF-kB 1999,Science, 285(5425): 260-3; activation, and/or co- Song HY et al., 1997Proc Natl Acad stimulation of immune Sci USA 94(18): 9792-6; Epsevikcells such as B and T and Nissen-Meyer, 1986, J. Immunol. cells.Methods. TNF-RII; TNF GeneSeq EP418014 Activities associated withapoptosis, Apoptosis activity, NF-kB activation, Soluble TNFR-II p75receptor; Accession R11141 NF-kB activation, and co-stimulation of and Band T cell co-stimulation can receptor polypeptides Death Receptorimmune cells such as T and B cells. be determined using assays known inmay be useful for the art: Moore et al., 1999, Science, inhibitingapoptosis, 285(5425): 260-3; Song HY et al., NF-kB activation, 1997 ProcNatl Acad Sci USA and/or co-stimulation of 94(18)9792-6; Epsevik andNissen- immune cells such as B Meyer, 1986, J. Immunol. Methods. and Tcells. hAPO-4; TROY GeneSeq WO9911791 Activities associated withapoptosis, Apoptosis activity, NF-kB activation, Immune Disorders,Accession NF-kB activation, and co-stimulation of and B and T cellco-stimulation can Cancers W93581 immune cells such as T and B cells. bedetermined using assays known in the art: Moore et al., 1999, Science,285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18):9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. TNF-alphaGeneSeq EP205038 Activities associated with apoptosis, Apoptosisactivity, NF-kB activation, Inflammatory disorders, precursor AccessionP60074 NF-kB activation, and co-stimulation of and B and T cellco-stimulation can immunologic disorders, immune cells such as T and Bcells. be determined using assays known in cancer the art: Moore et al.,1999, Science, 285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad SciUSA 94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol.Methods. Human TNF- GeneSeq EP619372 Activities associated withapoptosis, Apoptosis activity, NF-kB activation, Inflammatory disorders,alpha Accession R62463 NF-kB activation, and co-stimulation of and B andT cell co-stimulation can immunologic disorders, immune cells such as Tand B cells be determined using assays known in cancer the art: Moore etal., 1999, Science, 285(5425): 260-3; Song HY et al., 1997 Proc NatlAcad Sci USA 94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J.Immunol. Methods. Human TNF- GeneSeq EP563714 Activities associated withapoptosis, Apoptosis activity, NF-kB activation, Inflammatory disorders,alpha Accession R42679 NF-kB activation, and co-stimulation of and B andT cell co-stimulation can immunologic disorders, immune cells such as Tand B cells. be determined using assays known in cancer the art: Mooreet al., 1999, Science, 285(5425): 260-3; Song HY et al., 1997 Proc NatlAcad Sci USA 94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J.Immunol. Methods. Human TNF- GeneSeq WO0064479 Activities associatedwith apoptosis, Apoptosis activity, NF-kB activation, Inflammatorydisorders, beta (LT-alpha) Accession B37799 NF-kB activation, andco-stimulation of and B and T cell co-stimulation can immunologicdisorders, immune cells such as T and B cells. be determined usingassays known in cancer the art: Moore et al., 1999, Science, 285(5425):260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18): 9792-6;Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. LT-alpha GeneSeqEP250000 Activities associated with apoptosis, Apoptosis activity, NF-kBactivation, Inflammatory disorders, Accession P70107 NF-kB activation,and co-stimulation of and B and T cell co-stimulation can immunologicdisorders, immune cells such as T and B cells. be determined usingassays known in cancer the art: Moore et al., 1999, Science, 285(5425):260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18): 9792-6;Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. LT-beta GeneSeqWO9413808 Activities associated with apoptosis, Apoptosis activity,NF-kB activation, Inflammatory disorders, Accession R56869 NF-kBactivation, and co-stimulation of and B and T cell co-stimulation canimmunologic disorders, immune cells such as T and B cells. be determinedusing assays known in cancer the art: Moore et al., 1999, Science,285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad Sci USA94(18)9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. OPGLGeneSeq WO9846751 Activities associated with apoptosis, Apoptosisactivity, NF-kB activation, Inflammatory disorders, Accession NF-kBactivation, and co-stimulation of and B and T cell co-stimulation canimmunologic disorders, W83195 immune cells such as T and B cells. bedetermined using assays known in cancer, loss of bone the art: Moore etal., 1999, Science, mass 285(5425): 260-3; Song HY et al., 1997 ProcNatl Acad Sci USA 94(18)9792-6; Epsevik and Nissen- Meyer, 1986, J.Immunol. Methods. FasL GeneSeq WO9903999 Activities associated withapoptosis, Apoptosis activity, NF-kB activation, Inflammatory disorders,Accession NF-kB activation, and co-stimulation of and B and T cellco-stimulation can immunologic disorders, W98071 immune cells such as Tand B cells. be determined using assays known in cancer the art: Moore,et al., 1999, Science, 285(5425): 260-3; Song HY et al., 1997 Proc NatlAcad Sci USA 94(18)9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol.Methods. FasL GeneSeq WO9903998 Activities associated with apoptosis,Apoptosis activity, NF-kB activation, Inflammatory disorders, AccessionNF-kB activation, and co-stimulation of and B and T cell co-stimulationcan imunologic disorders, W95041 immune cells such as T and B cells. bedetermined using assays known in cancer the art: Moore et al., 1999,Science, 285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad Sci USA94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods.CD27L GeneSeq WO9405691 Activities associated with apoptosis, Apoptosisactivity, NF-kB activation, Inflammatory disorders, Accession R50121NF-kB activation, and co-stimulation of and B and T cell co-stimulationcan immunologic disorders, immune cells such as T and B cells. bedetermined using assays known in cancer the art: Moore et al., 1999,Science, 285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad Sci USA94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods.CD30 ligand GeneSeq WO9324135 Activities associated with apoptosis,Apoptosis activity, NF-kB activation, Inflammatory disorders, AccessionR45007 NF-kB activation, and co-stimulation of and B and T cellco-stimulation can immunologic disorders, immune cells such as T and Bcells. be determined using assays known in cancer the art: Moore et al.,1999, Science, 285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad SciUSA 94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol.Methods. CD40L GeneSeq WO9529935 Activities associated with apoptosis,Apoptosis activity, NF-kB activation, Inflammatory disorders, AccessionR85486 NF-kB activation, and co-stimulation of and B and T cellco-stimulation can immunologic disorders, immune cells such as T and Bcells. be determined using assays known in cancer the art: Moore, etal., 1999, Science, 285(5425): 260-3; Song HY et al., 1997 Proc NatlAcad Sci USA 94(18): 9792-6; Epsevik and Nissen- Meyer, 1986, J.Immunol. Methods. 4-1BB ligand GeneSeq U.S. Pat. No. 5,674,704Activities associated with apoptosis, Apoptosis activity, NF-kBactivation, Inflammatory disorders, Accession NF-kB activation, andco-stimulation of and B and T cell co-stimulation can immunologicdisorders, W26657 immune cells such as T and B cells. be determinedusing assays known in cancer the art: Moore et al., 1999, Science,285(5425): 260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18):9792-6; Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. FAS LigandGeneSeq WO0058465 Activities associated with apoptosis, Apoptosisactivity, NF-kB activation, Soluble DcR3 Inhibitory Accession B19335NF-kB activation, and co-stimulation of and B and T cell co-stimulationcan polypeptides may be Protein (DcR3) immune cells such as T and Bcells. be determined using assays known in useful for inhibiting theart: Moore et al., 1999, Science, apoptosis, NF-kB 285(5425): 260-3;Song HY et al., activation, and/or co- 1997 Proc Natl Acad Sci USAstimulation of immune 94(18): 9792-6; Epsevik and Nissen- cells such asB and T Meyer, 1986, J. Immunol. Methods cells. OX40L GeneSeq WO9521915Activities associated with apoptosis, Apoptosis activity, NF-kBactivation, Inflammatory disorders, Accession R79903 NF-kB activation,and co-stimulation of and B and T cell co-stimulation can immunologicdisorders, immune cells such as T and B cells. be determined usingassays known in cancer the art: Moore et al., 1999, Science, 285(5425):260-3; Song HY et al., 1997 Proc Natl Acad Sci USA 94(18): 9792-6;Epsevik and Nissen- Meyer, 1986, J. Immunol. Methods. Protease GeneSeqWO9106561 Peptides that inhibit the function/binding HIV proteaseactivities are known in HIV, inflammatory inhibitor Accessions of HIVthe art: HIV protease assays: disorders, immunologic peptides R12435,R12436, EP0387231. One can modify the disorders, cancer, viral R12437,R12438, assay to look for inhibition using any infections R12439,R12440, of the disclosed protease inhibitor and R1244 polypeptides.Retroviral protease GeneSeq EP387231 Peptides that inhibit thefunction/binding HIV protease activities are known in HIV, inflammatoryinhibitors Accessions of HIV the art: HIV protease assays: disorders,immunologic R06660, R06661, EP0387231. One can modify the disorders,cancer, viral R06662, R06663, assay to look for inhibition using anyinfections R06664, R06665, of the disclosed protease inhibitor R06666,R06667, polypeptides. R06668, R06669, R06670, R06671, R06672, R06673,R06674, R06675, and R06676 HIV protease GeneSeq WO9301828 Peptides thatinhibit the function/binding of HIV protease activities are known in theHIV, inflammatory inhibiting Accessions HIV art: HIV protease assays:EP0387231. disorders, immunologic peptides R59293, R59294, One canmodify, the assay to look for disorders, cancer, viral R59295, R59296,inhibition using any of the disclosed infections R59297, proteaseinhibitor polypeptides. R59298, R59299, R592300, R59301, R59302, R59301,R59302, R59303, R59304, R59305, R59306, R59307, R59308, R59309, R59310,R59311, R59312, R59313, R59314, R59315, R59316, R59317 R59318, R59319,R59320, R59321, R59322, R59323, R59324, R59325, R59326, R59327, R59328,R59329, R59330, R59331, R59332, R59333, R59334, R59335, R59336, R59337,R59338, R59339, R59340, R59341, R59342, R59343, R59344, R59345, R59346,R59347, R59348, R59349, and R59350 HIV-1 protease GeneSeq DE4412174Peptides that inhibit the function/binding of HIV protease activitiesare known in the HIV, inflammatory hinibitors Accessions HIV art: HIVprotease assays: EP0387231. disorders, immunologic R86326, R86327, Onecan modify the assay to look for disorders, cancer, viral R86328,R86329, inhibition using any of the disclosed infections R86330, R86331,protease inhibitor polypeptides. R86332, R86333, R86334, R86335, R86336,R86337, R86338, R86339, R86340, R86341, R86342, R86343, R86344, R86345,R86346, R86347, R86348, R86349, R86350, R86351, R86352, R86353, R86354,R86355, R86356, R86357, R86358, R86359, R86360, R86361, R86362, R86363,R86364, R86365, R86366, R86367, R86368, R86369, R86370, and R86371 HIVInhibitor GeneSeq WO9959615 Peptides that inhibit the function/bindingHIV protease activities are known in HIV, inflammatory Peptide Accessionof HIV the art: HIV protease assays: disorders, immunologic Y89687EP0387231. One can modify the disorders, cancer, viral assay to look forinhibition using any infections of the disclosed protease inhibitorpolypeptides. HIV Inhibitor GenSeq WO9948513 Peptides that inhibit thefunction/binding HIV Protease activities are known in HIV, inflammatoryPeptide Accession of HIV the art; HIV protease assays: disorders,immunologic Y31955 EP0387231. One can modify the disorders, cancer,viral assay to look for inhibition using any infections. of thedisclosed protease inhibitor polypeptides. HIV Inhibitorwww.sciencexpress.org; Peptides that inhibit the function/binding HIVprotease activities are known in HIV, inflammatory Peptide Published ofHIV the art: HIV protease assays: disorders, immunologic online 12 Jan.EP0387231: One can modify the disorders, cancer, viral 2001; assay tolook for inhibition using any infections 10.1126/science.1057453 of thedisclosed protease inhibitor polypeptides. Human monocyte GeneSeqWO9509232 Chemokines are a family of small, Chemokine activities can beImmune disorders, chemoattractant Accession secreted proteins involvedin biological determined using assays known in particularly useful forfactor hMCP-3 R73915 processes ranging from hematopoiesis, the art:Methods in Molecular treating bacterial and/or angiogenesis, andleukocyte trafficking. Biology, 2000, vol. 138: Chemokine viralmenigitis Members of this family are involved in a Protocols, Edited by:A. E. I. Proudfoot, similarly diverse range of pathologies T. N. C.Wells, and C. A. Power. including inflammation, allergy, tissue © HumanaPress Inc., rejection, viral infection, and tumor Totowa, NJ biology.The chemokines exert their effects by acting on a family of seventransmembrane G-protein-coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. Humanmonocyte GeneSeq WO9509232 Chemokines are a family of related Chemokineactivities can be Immune disorders, chemoattractant Accession small,secreted proteins involved in determined using assays known inparticularly useful for factor hMCP-1 R73914 biological processesranging from the art: Methods in Molecular treating bacterial and/orhematopoiesis, angiogenesis, and Biology, 2000, vol. 138: Chemokineviral menigitis leukocyte trafficking. Members of this Protocols. Editedby: A. E. I. Proudfoot, family are involved in a similarly T. N. C.Wells, and C. A. Power. diverse range of pathologies including © HumanaPress Inc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human gro-beta GeneSeq WO9429341Chemokines are a family of small, Chemokine activities can be immunedisorders, chemokine Accessions secreted proteins involved in biologicaldetermined using assays known in inflammatory disorders, R66699 andprocesses ranging from hematopoiesis, the art: Methods in Molecularblood-related disorders, W17671 angiogenesis, and leukocyte trafficking.Biology, 2000, vol. 138: Chemokine stem cell Members of this family areinvolved in a Protocols. Edited by: A. E. I. Proudfoot, transplantation,cancer similarly diverse range of pathologies T. N. C. Wells, and C. A.Power. including inflammation, allergy, tissue © Humana Press Inc.,rejection, viral infection, and tumor Totowa, NJ. biology. Thechemokines exert their effects by acting on a family of seventransmembrane G-protein-coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. Humangro- GeneSeq WO9429341 Chemokines are a family of related Chemokineactivities can be Immune disorders, gamma chemokine Accessions small,secreted proteins involved in determined using assays known ininflammatory disorders, R66700 and biological processes ranging from theart: Methods in Molecular blood-related disorders, W17672 hematopoiesis,angiogenesis, and Biology, 2000, vol. 138: Chemokine stem cell leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,transplantation, cancer family are involved in a similarly T. N. C.Wells, and C. A. Power. diverse range of pathologies including © HumanaPress Inc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified Human gro-alpha GeneSeq WO9429341Chemokines are a family of related Chemokine activities can be Immunedisorders, chemokine Accessions small, secreted proteins involved indetermined using assays known in inflammatory disorders, R66698 andbiological processes ranging from the art: Methods in Molecularblood-related disorders, W18024 hematopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine stem cell leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot,transplantation, cancer family are involved in a similarly T. N. C.Wells, and C. A. Power. diverse range of pathologies including © HumanaPress Inc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified Human eosinophil- GeneSeq WO9632481Chemokines are a family of related Chemokine activities can be Immunedisorders, expressed Accession small, secreted proteins involved indetermined using assays known in particularly treatment of chemokine(EEC) W05186 biological processes ranging from the art: Methods inMolecular eosinophilia, hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine inflammation, allergies, leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, asthma,leukaemia and family are involved in a similarly T. N. C. Wells, and C.A. Power. lymphoma diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified Chemokine-like GeneSeq WO9613587Chemokines are a family of related Chemokine activities can be Cancerand blood- protein PF4-414 Accessions small, secreted proteins involvedin determined using assays known in related disorders, Full-Length andR92318 and biological processes ranging from the art: Methods inMolecular particularly Mature R99809 hematopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine myelosuppression leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Power.diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified Chemokine-like GeneSeq WO9613587 Chemokines are a family ofrelated Chemokine activities can be Cancer and blood- protein IL-8M3Accession small, secreted proteins involved in determined using assaysknown in related disorders, R99812 biological processes ranging from theart: Methods in Molecular particularly hematopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine myelosuppression leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Power.diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ; and Holmes et al(1991) viral infection, and tumor biology. The Science 253, 1278-80.chemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identifiedHuman GeneSeq WO9613587 Chemokines are a family of related Chemokineactivities can be Cancer and blood- interleukin-8 (IL- Accession small,secreted proteins involved in determined using assays known in relateddisorders, 8) R99814 biological processes ranging from the art: Methodsin Molecular particularly hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine myelosuppression leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power. diverse rangeof pathologies including © Humana Press Inc., inflammation, allergy,tissue rejection, Totowa, NJ; and Holmes et al (1991) viral infection,and tumor biology. The Science 253, 1278-80. chemokines exert theireffects by acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified Chemokine-like GeneSeq WO9613587Chemokines are a family of related Chemokine activities can be Cancerand blood- protein IL-8M1 Accessions small, secreted proteins involvedin determined using assays known in related disorders, Full-Length andR99815 and biological processes ranging from the art: Methods inMolecular particularly Mature R99803 hematopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine myelosuppression leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Power.diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ; and Holmes et al(1991) viral infection, and tumor biology. The Science 253, 1278-80.chemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identifiedChemokine-like GeneSeq WO9613587 Chemokines are a family of relatedChemokine activities can be Cancer and blood- protein IL-8M8 Accessionssmall, secreted proteins involved in determined using assasys known inrelated disorders, Full-Length and R99816 and biological processesranging from the art: Methods in Molecular particularly Mature R99805hematopoiesis, angiogenesis, and Biology, 2000, vol. 138: Chemokinemyelosuppression. leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa,NJ; and Holmes et al (1991) viral infection, and tumor biology. TheScience 253, 1278-80. chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Chemokine-like GeneSeq WO9613587 Chemokines are a familyof related Chemokine activities can be Cancer and blood- protein IL-8M8Accessions small, secreted proteins involved in determined using assasysknown in related disorders, Full-Length and R99817 and biologicalprocesses ranging from the art: Methods in Molecular particularly MatureR99806 hematopoiesis, angiogenesis, and Biology, 2000, vol. 138:Chemokine myelosuppression. leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot; family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ; and Holmes et al (1991) viral infection, and tumor biology.The Science 253, 1278-80. chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Chemokine-like GeneSeq WO9613587 Chemokines are a familyof related Chemokine activities can be Cancer and blood- protein IL-8M8Accessions small, secreted proteins involved in determined using assasysknown in related disorders, Full-Length and R99818 and biologicalprocesses ranging from the art: Methods in Molecular particularly MatureR99804 hematopoiesis, angiogenesis, and Biology, 2000, vol. 138:Chemokine myelosuppression. leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot; family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ; and Holmes et al (1991) viral infection, and tumor biology.The Science 253, 1278-80. chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Chemokine-like GeneSeq WO9613587 Chemokines are a familyof related Chemokine activities can be Cancer and blood- protein IL-8M8Accessions small, secreted proteins involved in determined using assasysknown in related disorders, Full-Length and R99819 and biologicalprocesses ranging from the art: Methods in Molecular particularly MatureR99807 hematopoiesis, angiogenesis, and Biology, 2000, vol. 138:Chemokine myelosuppression. leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot; family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Chemokine-like GeneSeqWO9613587 Chemokines are a family of related Chemokine activities can beCancer and blood- protein IL-8M8 Accessions small, secreted proteinsinvolved in determined using assasys known in related disorders,Full-Length and R99822 and R9807 biological processes ranging from theart: Methods in Molecular particularly Mature hematopoiesis,angiogenesis, and Biology, 2000, vol. 138: Chemokine myelosuppression.leukocyte trafficking. Members of this Protocols. Edited by: A. E. I.Proudfoot; family are involved in a similarly T. N. C. Wells, and C. A.Power. diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human foetal GeneSeq WO9622374 Chemokines are a family ofrelated Chemokine activities can be Immune disorders spleen expressedAccession R98499 small, secreted proteins involved in determined usingassasys known in chemokine, FSEC biological processes ranging from theart: Methods in Molecular hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot; family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Liver expressed GeneSeqWO9616979 Chemokines are a family of related Chemokine activities can beInflammation of the chemokine- Accession R95689 small, secreted proteinsinvolved in determined using assasys known in liver 1(LVEC-1) biologicalprocesses ranging from the art: Methods in Molecular hematopoiesis,angiogenesis, and Biology, 2000, vol. 138: Chemokine leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot;family are involved in a similarly T. N. C. Wells, and C. A. Power.diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Liver expressed GeneSeq WO9616979 Chemokines are a family ofrelated Chemokine activities can be Inflammation of the chemokine-Accession R95690 small, secreted proteins involved in determined usingassasys known in liver 2(LVEC-2) biological processes ranging from theart: Methods in Molecular hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot; family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Pituitary expressedGeneSeq WO9616979 Chemokines are a family of related Chemokineactivities can be Inflammation, chemokine Accession R95691 small,secreted proteins involved in determined using assasys known inparticularly of the liver (PGEC) biological processes ranging from theart: Methods in Molecular hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot; family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines liave beendescribed, which bind to ~17 receptors thus far identified.Adenoid-expressed GeneSeq WO9617868 Chemokines are a family of relatedChemokine activities can be Inflammation, chemokine Accession R97664small, secreted proteins involved in determined using assasys known inangiogenesis, (ADEC) biological processes ranging from the art: Methodsin Molecular tumorigenesis, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine musculoskeletal leukocyte trafficking. Membersof this Protocols. Edited by: A. E. I. Proudfoot; disorders family areinvolved in a similarly T. N. C. Wells, and C. A. Power. diverse rangeof pathologies including © Humana Press Inc., inflammation, allergy,tissue rejection, Totowa, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Human GeneSeq Chemokines are a family of related Chemokine activitiescan be Immune disorders, cell chemokineCC-2 Accession small, secretedproteins involved in determined using assays known in migration,proliferation, W38170 biological processes ranging from the art: Methodsin Molecular and differentiation, hematopoiesis, angiogenesis, andBiology, 2000, vol. 138; Chemokine disorders leukocyte trafficking.Members of this protocols. Edited by: A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power. diverse rangeof pathologies including © Humana Press Inc. inflammation, allergy,tissue rejection, Totowa, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G-protein-coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. HumanGeneSeq WO9741230 Chemokines are a family of related Chemokineactivities can be Immune disorders, cell chemokine Accession small,secreted proteins involved in determined using assays known in themigration, proliferation, HCC-1 W38171 biological processes ranging fromart: Methods in molecular Biology and differentiation hematopiesis,anglogenesis and 2000, vol. 138: Chemokine disorders leukocytetrafficking. Members of this Protocols. Edited by A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells and C. A. Powerdiverse range of pathologies including © Humana Press Inc., Totowa,inflammation, allergy, tissue rejection, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G-protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human GeneSeq WO9741230 Chemokines are a family of relatedChemokine activities can be Immune disorders, cell chemokine CC-3Accession small, secreted proteins involved in determined using assaysknown in the migration, proliferation W38172 biological processesranging from art: Methods in molecular Biology, and differentiationhemotopoiesis, anglogenesis, and 2000, vol. 138: Chemokine disordersleukocyte trafficking. Members of this Protocols, Edited by A. E. I.Proudfoot, family are involved in a similarly T. N. C. Wells, and C. A.Power diverse range of pathologies including © Humana Press Inc.,Totowa, inflammation, allergy, tissue rejection, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Novel GeneSeq WO9739126 Chemokines are a family of relatedChemokine activities can be Immune disorders, betachemokine Accessionsmall, secreted proteins involved in determined using assays known inthe vascular disorders, designated PTEC W27271 biological processesranging from art: Methods in molecular Biology, cancer hemotopoiesis,anglogenesis, and 2000, vol. 138: Chemokine leukocyte trafficking.Members of this Protocols, Edited by A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power diverse range ofpathologies including © Humana Press Inc., Totowa, inflammation,allergy, tissue rejection, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Human CX3C GeneSeq WO9727299 Chemokines are a family of relatedChemokine activities can be Immune disorders, 111 amino acid Accessionsmall, secreted proteins involved in determined using assays known inthe inflammatory diseases, chemokine W23344 biological processes rangingfrom art: Methods in molecular Biology, abnormal proliferation,hemotopoiesis, anglogenesis, and 2000, vol. 138: Chemokine regeneration,leukocyte trafficking. Members of this Protocols, Edited by A. E. I.Proudfoot, degeneration, and family are involved in a similarly T. N. C.Wells, and C. A. Power atrophy diverse range of pathologies including© Humana Press Inc., Totowa, inflammation, allergy, tissue rejection, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human CCF18 GeneSeq WO9721812Chemokines are a family of related Chemokine activities can be Abnormalphysiology chemokine Accession small, secreted proteins involved indetermined using assays known in the and development W25942 biologicalprocesses ranging from art: Methods in molecular Biology, disorders, canalso be hemotopoiesis, anglogenesis, and 2000, vol. 138: Chemokine usedas an anti-viral leukocyte trafficking. Members of this Protocols,Edited by A. E. I. Proudfoot, agent family are involved in a similarlyT. N. C. Wells, and C. A. Power diverse range of pathologies including© Humana Press Inc., Totowa, inflammation, allergy, tissue rejection, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human beta- GeneSeq WO9725427Chemokines are a family of related Chemokine activities can beChemotaxis, chemokine Accession small, secreted proteins involved indetermined using assays known in the blood-related disorders, H1305(MCP-2) W26655 biological processes ranging from art: Methods inmolecular Biology, viral infection, HIV, hemotopoiesis, anglogenesis,and 2000, vol. 138: Chemokine wound healing, cancer leukocytetrafficking. Members of this Protocols, Edited by A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Powerdiverse range of pathologies including © Humana Press Inc., Totowa,inflammation, allergy, tissue rejection, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human GeneSeq WO9712914 Chemokines are a family of relatedChemokine activities can be Inflammatory and eosinocyte CC Accessionsmall, secreted proteins involved in determined using assays known inthe immune disorders type chemokine W14990 biological processes rangingfrom art: Methods in molecular Biology, eotaxin hemotopoiesis,anglogenesis, and 2000, vol. 138: Chemokine leukocyte trafficking.Members of this Protocols, Edited by A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power diverse range ofpathologies including © Humana Press Inc., Totowa, inflammation,allergy, tissue rejection, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Human thymus GeneSeq WO9711969 Chemokines are a family of relatedChemokine activities can be Inflammatory and and activation Accessionsmall, secreted proteins involved in determined using assays known inthe immune disorders regulated W14018 biological processes ranging fromart: Methods in molecular Biology, cytokine hemotopoiesis, anglogenesis,and 2000, vol. 138: Chemokine (TARC) leukocyte trafficking. Members ofthis Protocols, Edited by A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power diverse range of pathologiesincluding © Humana Press Inc., Totowa, inflammation, allergy, tissuerejection, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human GeneSeq WO9712041Chemokines are a family of related Chemokine activities can be Cancer,would healing, chemokine beta- Accession small, secreted proteinsinvolved in determined using assays known in the immune disorders 8short forms W16315 biological processes ranging from art: Methods inmolecular Biology, hemotopoiesis, anglogenesis, and 2000, vol. 138:Chemokine leukocyte trafficking. Members of this Protocols, Edited by A.E. I. Proudfoot, family are involved in a similarly T. N. C. Wells, andC. A. Power diverse range of pathologies including © Humana Press Inc.,Totowa, inflammation, allergy, tissue rejection, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Microphage GeneSeq WO9640923 Chemokines are a family ofrelated Chemokine activities can be Inflammatory derived Accessionsmall, secreted proteins involved in determined using assays known indiseases, wound chemokine, MDC W20058 biological processes ranging fromthe art: Methods in Molecular healin, hermaatopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine angiogenesis leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power. diverse rangeof pathologies including © Humana Press Inc., inflammation, allergy,tissue rejection, Totowa, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Human chemokine GeneSeq WO9844117 Chemokines are a family of relatedChemokine activities can be Inflammatory and ZSIG-35 Accession small,secreted proteins involved in determined using assays known in immunediseases W30565 biological processes ranging from the art: Methods inMolecular hematopoiesis, angiogenesis, and Biology, 2000, vol. 138:Chemokine leukocyte trafficking. Members of this Protocols. Edited by:A. E. I. Proudfoot, family are involved in a similarly T. N. C. Wells,and C. A. Power. diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Primate CC GeneSeq WO98328658 Chemokinesare a family of related Chemokine activities can be Immune and chemokineAccesssion small, secreted proteins involved in determined using assaysknown in inflammatory “ILINCK” W69990 biological processes ranging fromthe art: Methods in Molecular disorders, hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine abnormal leukocyte trafficking.Protocols. Edited by: A. E. I. Prodfoot, proliferation, T. N. C. Wells,and C. A. Power. regeneration, © Humana Press Inc., generation andTotowa, NJ atrophy disorders Primate CXC GeneSeq WO9832858 Chemokinesare a family of related Chemokine activities can be Immune and chemokineAccession small, secreted proteins involved in determined using assaysknown in inflammatory “IBICK” W69989 biological processes ranging fromthe art: Methods in Molecular disorders, hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine abnormal leukocyte trafficking.Members of this Protocols. Editd by: A. E. I. Proudfoot, proliferation,family are involved in a similarly T. N. C. Wells, and C. A. Power.regeneration, diverse range of pathologies including © Humana PressInc., generation and inflammation, allergy, tissue rejection, Totowa, NJatrophy disorders viral infection, and tumor biology. The chemokinesexert their effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human CC-type GeneSeqWO9831809 Chemokines are a family of related Chemokine activities can beImmune, chemokine protein Accession small, secreted proteins involved indetermined using assays known in inflammatory, and designated SLC W69163biological processes ranging from the art: Methods in Molecularinfectious (secondary hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine disorders, cancer lymphoid leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, chemokine)family are involved in a similarly T. N. C. Wells, and C. A. Power.diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human CC GeneSeq WO9826071 Chemokines are a family ofrelated Chemokine activities can be Cancer and chemokine ELC Accessionsmall, secreted proteins involved in determined using assays known ininfectious protein W62542 biological processes ranging from the art:Methods in Molecular diseases, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine particularly leukocyte trafficking. Members ofthis Protocols. Edited by: A. E. I. Proudfoot, herpes virus family areinvolved in a similarly T. N. C. Wells, and C. A. Power. diverse rangeof pathologies including © Humana Press Inc., inflammation, allergy,tissue rejection, Totowa, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Human DVic-1 GeneSeq Wo9823750 Chemokines are a family of relatedChemokine activities can be Abnormal C-C chemokine Accession small,secreted proteins involved in determined using assays known inproliferation, W60649 biological processes ranging from the art: Methodsin Molecular regeneration, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine degeneration, and leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, atrophydisorders, family are involved in a similarly T. N. C. Wells, and C. A.Power. including cancer diverse range of pathologies including © HumanaPress Inc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human C-C GeneSeq WO9823750 Chemokinesare a family of related Chemokine activities can be Immune chemokineAccession small, secreted proteins involved in determined using assaysknown in disorders, cell DGWCC W60650 biological processes ranging fromthe art: Methods in Molecular proliferation hematophoiesis,angiogenesis, and Biology, 2000, vol. 138: Chemokine disorders, cancerleukocyte trafficking. Members of this Protocols. Edited by: A. E. I.Proudfoot, family are involved in a similarly T. N. C. Wells, and C. A.Power. diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentifed. Human STCP-1 GeneSeq WO9824907 Chemokines are a family ofrelated Chemokine activities can be Immune Accession small, secretedproteins involved in determined using assays known in disorders, W62783biological processes ranging from the art: Methods in Molecularparticularly T cell hematopoiesis, angiogenesis, and Biology, 2000, vol.138: Chemokine related disorders, leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, viral infection, family areinvolved in a similarly T. N. C. Wells, and C. A. Power. andinflammation, diverse range of pathologies including © Humana PressInc., especially joint inflammation, allergy, tissue rejection, Totowa,NJ viral infection, and tumor biology. The chemokines exert theireffects by acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Exodua protein GeneSeq WO9821330Chamokines are a family of related Chemokine activities can be Immuneand Accession small, secreted proteins involved in determined usingassays known in inflammatory W61279 biological processes ranging fromthe art: Methods in Molecular disorders, hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine angiogenesis, leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,cancer, and family are involved in a similarly T. N. C. Wells, and C. A.Power. proliferation diverse range of pathologies including © HumanaPress Inc., disorders, inflammation, allergy, tissue rejection, Totowa,NJ particularly viral infection, and tumor biology. Themyeloproliferative chemokines exert their effects by acting diseases ona family of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human GeneSeq WO9814581 Chemokines are a family ofrelated Chemokine activities can be Cancer and degenerative Chr19KineAcession small, secreted proteins involved in determined using assaysknown in disorders protein W50887 biological processes ranging from theart: Methods in Molecular hematopoiesis, angiogenesis, and Biology,2000, vol. 138: leukocyte trafficking. Members of this ChemokineProtocols, Edited by: family are involved in a similarly A. E. I.Proudfoot, T. N. C. Wells, and diverse range of pathologies including C.A. Power. © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human T cell GeneSeqU.S. Pat. No. 5,780,268 Chemokines area family of related Chemokineactivities can be Immune, inflammatory, mixed Accession small, secretedproteins involved in determined using assays known in the and infectiousdisorders, lymphocyte W58703 biological processes ranging from art:Mehtods of Molecular Biology, cancer reaction hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine expressed leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,chemokine family are involved in a similarly T. N. C. Wells, and C. A.Power (TMEC) diverse range of pathologies including © Humana Press Inc.,Totowa, inflammation, allergy, tissue rejection, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human 6CKine GeneSeq W09814581 Chemokines area family ofrelated Chemokine activities can be Cancer and degenerative proteinAccession small, secreted proteins involved in determined using assaysknown in the disorders W50885 biological processes ranging from art:Mehtods of Molecular Biology, hematopoiesis, angiogenesis, and 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power diverse range of pathologies including© Humana Press Inc., Totowa, inflammation, allergy, tissue rejection, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. human liver and GeneSeq WO9817800Chemokines area family of related Chemokine activities can be Immune,inflammatory, activation Accession small, secreted proteins involved indetermined using assays known in the and infectious disorders, regulatedW57475 biological processes ranging from art: Mehtods of MolecularBiology, cancer chemokine hematopoiesis, angiogenesis, and 2000, vol.138: Chemokine (LARC) leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power diverse range of pathologies including© Humana Press Inc., Totowa, inflammation, allergy, tissue rejection, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. RANTES GeneSeq WO9744462 Chemokinesarea family of related Chemokine activities can be Infectious diseases,peptide Accession small, secreted proteins involved in determined usingassays known in the particularly HIV W29538 biological processes rangingfrom art: Mehtods of Molecular Biology, hematopoiesis, angiogenesis, and2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power diverse range of pathologiesincluding © Humana Press Inc., Totowa, inflammation, allergy, tissuerejection, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. RANTES 8-68 GeneSeqWO9744462 Chemokines area family of related Chemokine activities can beInfectious diseases, Accession small, secreted proteins involved indetermined using assays known in the particularly HIV W29529 biologicalprocesses ranging from art: Mehtods of Molecular Biology, hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power diverse range ofpathologies including © Humana Press Inc., Totowa, inflammation,allergy, tissue rejection, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.RANTES 9-68 GeneSeq WO9744462 Chemokines area family of relatedChemokine activities can be Infectious diseases, Accession small,secreted proteins involved in determined using assays known in theparticularly HIV W29528 biological processes ranging from art: Mehtodsof Molecular Biology, hematopoiesis, angiogenesis, and 2000, vol. 138:Chemokine leukocyte trafficking. Members of this Protocols. Edited by:A. E. I. Proudfoot, family are involved in a similarly T. N. C. Wells,and C. A. Power diverse range of pathologies including © Humana PressInc., Totowa, inflammation, allergy, tissue rejection, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human GeneSeq WO9811226 Chemokines areafamily of related Chemokine activities can be Abnormal proliferation,chemokine Accession small, secreted proteins involved in determinedusing assays known in the regeneration, protein 331D5 W59433 biologicalprocesses ranging from art: Mehtods of Molecular Biology, degenerationor atrophy, hematopoiesis, angiogenesis, and 2000, vol. 138: Chemokineincluding cancer leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power diverse range of pathologies including© Humana Press Inc., Totowa, inflammation, allergy, tissue rejection, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human GeneSeq WO9811226 Chemokinesarea family of related Chemokine activities can be Abnormalproliferation, chemokine Accession small, secreted proteins involved indetermined using assays known in the regeneration, protein 61164 W59430biological processes ranging from art: Mehtods of Molecular Biology,degeneration or atrophy, hematopoiesis, angiogenesis, and 2000, vol.138: Chemokine including cancer leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power diverse range of pathologiesincluding © Humana Press Inc., Totowa, inflammation, allergy, tissuerejection, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Chemokine GeneSeqWO9809171 Chemokines area family of related Chemokine activities can beImmune, Inflammatory, MCP-4 Accession small, secreted proteins involvedin determined using assays known in the and infectious diseases W56690biological processes ranging from art: Mehtods of Molecular Biology,hematopoiesis, angiogenesis, and 2000, vol. 138: Chemokine leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Powerdiverse range of pathologies including © Humana Press Inc., Totowa,inflammation, allergy, tissue rejection, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human stromal GeneSeq FR2751658 Chemokines are a family ofrelated Chemokine activities can be HIV infections cell-derivedAccession small, secreted proteins involved in determined using assaysknown in chemokine, SDF-1 W50766 biological processes ranging from theart: Methods in Molecular hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Thymus expressedGeneSeq WO9801557 Chemokines are a family of related Chemokineactivities can be Immune and chemokine Accession small, secretedproteins involved in determined using assays known in the inflammatorydisorders (TECK) W44397 biological processes ranging from art: Methodsin Molecular Biology, hematopoiesis, angiogenesis, and 2000, vol. 138:Chemokine Protocols. leukocyte trafficking. Members of this Edited by:A. E. I. Proudfoot, T. N. C. Wells, family are involved in a similarlyand C. A. Power. © Humana diverse range of pathologies including PressInc., Totowa, NJ inflammation, allergy, tissue rejection, viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human chemokine GeneSeq WO9801557Chemokines are a family of related Chemokine activities can be Immuneand MIP-3alpha Accession small, secreted proteins involved in determinedusing assays known in the inflammatory disorders W44398 biologicalprocesses ranging from art: Methods in Molecular Biology, hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine Protocols. leukocytetrafficking. Members of this Edited by: A. E. I. Proudfoot, T. N. C.Wells, family are involved in a similarly and C. A. Power. © Humanadiverse range of pathologies including Press Inc., Totowa, NJinflammation, allergy, tissue rejection, viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human chemokine GeneSeq WO9801557 Chemokines are a family ofrelated Chemokine activities can be Immune and MIP-3beta Accessionsmall, secreted proteins involved in determined using assays known inthe inflammatory disorders W44399 biological processes ranging from art:Methods in Molecular Biology, hematopoiesis, angiogenesis, and 2000,vol. 138: Chemokine Protocols. leukocyte trafficking. Members of thisEdited by: A. E. I. Proudfoot, T. N. C. Wells, family are involved in asimilarly and C. A. Power. © Humana diverse range of pathologiesincluding Press Inc., Totowa, NJ inflammation, allergy, tissuerejection, viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human monocyte GeneSeqWO9802459 Chemokines are a family of related Chemokine activities can beImmune disorders, chemotactic Accession small, secreted proteinsinvolved in determined using assays known in the respiratory disorders,proprotein (MCPP) W42072 biological processes ranging from art: Methodsin Molecular Biology, cancer sequence hematopoiesis, angiogenesis, and2000, vol. 138: Chemokine Protocols. leukocyte trafficking. Members ofthis Edited by: A. E. I. Proudfoot, T. N. C. Wells, family are involvedin a similarly and C. A. Power. © Humana diverse range of pathologiesincluding Press Inc., Totowa, NJ inflammation, allergy, tissuerejection, viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Macrophage- GeneSeqU.S. Pat. No. 5,688,927/ Chemokines are a family of related Chemokineactivities can be Immune, and derived chemokine Accessions U.S. Pat. No.5,932,703 small, secreted proteins involved in determined using assaysknown in the inflammatory disorders, (MDC) W40811 and biologicalprocesses ranging from art: Methods in Molecular Biology, cancer Y24414hematopoiesis, angiogenesis, and 2000, vol. 138: Chemokine Protocols.leukocyte trafficking. Members of this Edited by: A. E. I. Proudfoot, T.N. C. Wells, family are involved in a similarly and C. A. Power.© Humana diverse range of pathologies including Press Inc., Totowa, NJinflammation, allergy, tissue rejection, viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Macrophage GeneSeq U.S. Pat. No. 5,932,703 Chemokines are afamily of related Chemokine activities can be Immune and derivedchemokine Accession Y24416 small, secreted proteins involved indetermined using assays known in the inflammatory disorders analogueMDC- biological processes ranging from art: Methods in MolecularBiology, eyfy hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. leukocyte trafficking. Members of this Edited by: A. E. I.Proudfoot, T. N. C. Wells, family are involved in a similarly and C. A.Power. © Humana diverse range of pathologies including Press Inc.,Totowa, NJ inflammation, allergy, tissue rejection, viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Macrophage GeneSeq U.S. Pat. No. 5,932,703 Chemokines are afamily of related Chemokine activities can be Immune and derivedchemokine Accession Y24413 small, secreted proteins involved indetermined using assays known in the inflammatory disorders analogue MDCbiological processes ranging from art: Methods in Molecular Biology,(n + 1) hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. leukocyte trafficking. Members of this Edited by: A. E. I.Proudfoot, T. N. C. Wells, family are involved in a similarly and C. A.Power. © Humana diverse range of pathologies including Press Inc.,Totowa, NJ inflammation, allergy, tissue rejection, viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Macrophage GeneSeq U.S. Pat. No. 5,932,703 Chemokines are afamily of related Chemokine activities can be Immune and derivedchemokine Accession Y24415 small, secreted proteins involved indetermined using assays known in the inflammatory disorders analogueMDC-yl biological processes ranging from art: Methods in MolecularBiology, hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. leukocyte trafficking. Members of this Edited by: A. E. I.Proudfoot, T. N. C. Wells, family are involved in a similarly and C. A.Power. © Humana diverse range of pathologies including Press Inc.,Totowa, NJ inflammation, allergy, tissue rejection, viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human type CC GeneSeq JP11243960 Chemokines are a family ofrelated Chemokine activities can be Allergic diseases and chemokineeotaxin Accession Y43178 small, secreted proteins involved in determinedusing assays known in the HIV infection 3 protein sequence biologicalprocesses ranging from art: Methods in Molecular Biology, hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine Protocols. leukocytetrafficking. Members of this Edited by: A. E. I. Proudfoot, T. N. C.Wells, family are involved in a similarly and C. A. Power. © Humanadiverse range of pathologies including Press Inc., Totowa, NJinflammation, allergy, tissue rejection, viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human MCP-3 GeneSeq WO9946392 Chemokines are a family ofrelated Chemokine activities can be Cancer and immune and human Muc-1Acession Y29893 small, secreted proteins involved in determined usingassays known in disorders, particularly core epitope biologicalprocesses ranging from the art: Methods in Molecular HIV infection (VNT)fusion hematopoiesis, angiogenesis, and Biology, 2000, vol. 138:Chemokine protein leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarily T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human IP-10 and GeneSeq WO9946392Chemokines are a family of related Chemokine activities can be Cancerand immune human Muc-1 core Accession Y29894 small, secreted proteinsinvolved in determined using assays known in the disorders, particularlyepitope (VNT) biological processes ranging from art: Methods inMolecular Biology, HIV infection fusion protein hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine Protocols. leukocytetrafficking. Members of this Edited by: A. E. I. Proudfoot, T. N. C.Wells, family are involved in a similarily and C. A. Power. © Humanadiverse range of pathologies including Press Inc., Totowa, NJinflammation, allergy, tissue rejection, viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human IP-10 and GeneSeq W09946392 Chemokines are a family ofrelated Chemokine activities can be Cancer and immune HIV-1 gp 120Accession Y29897 small, secreted proteins involved in determined usingassays known in the disorders, particularly hypervariable biologicalprocesses ranging from art: Methods in Molecular Biology, HIV infectionregion fusion hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. protein leukocyte trafficking. Members of this Edited by: A.E. I. Proudfoot, T. N. C. Wells, family are involved in a similarily andC. A. Power. © Humana diverse range of pathologies including Press Inc.,Totowa, NJ inflammation, allergy, tissue rejection, viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human mammary GeneSeq WO9936540 Chemokines are a family ofrelated Chemokine activities can be Breast disease, associatedAccessions small, secreted proteins involved in determined using assaysknown in the including cancer chemokine Y29092 and biological processesranging from art: Methods in Molecular Biology, (MACK) protein Y29093hematopoiesis, angiogenesis, and 2000, vol. 138: Chemokine Protocols.Full-Length and leukocyte trafficking. Members of this Edited by: A. E.I. Proudfoot, T. N. C. Wells, Mature family are involved in a similarilyand C. A. Power. © Humana diverse range of pathologies including PressInc., Totowa, NJ inflammation, allergy, tissue rejection, viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Tim-1 protein GeneSeq WO9933990Chemokines are a family of related Chemokine activities can beInflammation due to Accession small, secreted proteins involved indetermined using assays known in the stimuli such as heart Y28290biological processes ranging from art: Methods in Molecular Biology,attacks and stroke, hematopoiesis, angiogenesis, and 2000, vol. 138:Chemokine Protocols. infection, physical leukocyte trafficking. Membersof this Edited by: A. E. I. Proudfoot, T. N. C. Wells, trauma, UV orionizing family are involved in a similarily and C. A. Power. © Humanaradiation, burns, diverse range of pathologies including Press Inc.,Totowa, NJ frostbite or corrosive inflammation, allergy, tissuerejection, chemicals viral infection, and tumor biology. The chemokinesexert their effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human Lkn-1 GeneSeqWO9928473 and Chemokines are a family of related Chemokine activitiescan be HIV infection and Full-Length and Accessions WO9928472 small,secreted proteins involved in determined using assays known in thecancer, particularly Mature protein Y17280, Y17274, biological processesranging from art: Methods in Molecular Biology, leukemia Y17281, andhematopoiesis, angiogenesis, and 2000, vol. 138: Chemokine Protocols.Y17275 leukocyte trafficking. Members of this Edited by: A. E. I.Proudfoot, T. N. C. Wells, family are involved in a similarily and C. A.Power. © Humana diverse range of pathologies including Press Inc.,Totowa, NJ inflammation, allergy, tissue rejection, viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. N-terminal GeneSeq WO9920759 Chemokines are a family ofrelated Chemokine activities can be Inhibit or stimulate modifiedAccession Y05818 small, secreted proteins involved in determined usingassays known in the angiogenesis, inhibit the chemokine met- biologicalprocesses ranging from art: Methods in Molecular Biology, binding of HIVhSDF-1 alpha hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. leukocyte trafficking. Members of this Edited by: A. E. I.Proudfoot, T. N. C. Wells, family are involved in a similarily and C. A.Power. © Humana diverse range of pathologies including Press Inc.,Totowa, NJ inflammation, allergy, tissue rejection, viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. N-terminal GeneSeq WO9920759 Chemokines are a family ofrelated Chemokine activities can be Inhibit or stimulate modifiedAccession Y05819 small, secreted proteins involved in determined usingassays known in the angiogenesis, inhibit the chemokine met- biologicalprocesses ranging from art: Methods in Molecular Biology, binding ofHIV, hSDF-1 beta hematopoiesis, angiogenesis, and 2000, vol. 138:Chemokine Protocols. antiinflammatory; leukocyte trafficking. Members ofthis Edited by: A. E. I. Proudfoot, T. N. C. Wells, immunosuppressantfamily are involved in a similarily and C. A. Power. © Humana diverserange of pathologies including Press Inc., Totowa, NJ inflammation,allergy, tissue rejection, viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.N-terminal GeneSeq WO9920759 Chemokines are a family of relatedChemokine activities can be Inhibit or stimulate modified AccessionY05820 small, secreted proteins involved in determined using assaysknown in the angiogenesis, inhibit the chemokine biological processesranging from art: Methods in Molecular Biology, binding of HIV,GroHEK/hSDF- hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. antiinflammatory; 1alpha leukocyte trafficking. Members ofthis Edited by: A. E. I. Proudfoot, T. N. C. Wells, immunosuppressantfamily are involved in a similarily and C. A. Power. © Humana diverserange of pathologies including Press Inc., Totowa, NJ inflammation,allergy, tissue rejection, viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.N-terminal GeneSeq WO9920759 Chemokines are a family of relatedChemokine activities can be Inhibit or stimulate modified AccessionY05821 small, secreted proteins involved in determined using assaysknown in the angiogenesis, inhibit the chemokine biological processesranging from art: Methods in Molecular Biology, binding of HIV,GroHEK/hSDF- hematopoiesis, angiogenesis, and 2000, vol. 138: ChemokineProtocols. antiinflammatory; 1beta. leukocyte trafficking. Members ofthis Edited by: A. E. I. Proudfoot, T. N. C. Wells, immunosuppressantfamily are involved in a similarily and C. A. Power. © Humana diverserange of pathologies including Press Inc., Totowa, NJ inflammation,allergy, tissue rejection, viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Chemokine GeneSeq WO9912968 Chemokines are a family of related Chemokineactivities can be Increase or enhance an Eotaxin Accession small,secreted proteins involved in determined using assays known ininflammatory response, Y14230 biological processes ranging from the art:Methods in Molecular an immune response hematopoiesis, agiogenesis, andBilogy, 2000, vol. 138: Chemokine orhaematopoietic cell- leukocyetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,associated activity; treat family are involved in a similarly T. N. C.Wells, and C. A. Power. a vascular indication; diverse range ofpathologies including © Humana Press Inc., Cancer; enhance woundinflammation, allergy, tissue rejection, Totowa, NJ healing, to preventor viralk infection, and tumor biology. The treat asthma, organchemokines exert their effects by acting transplant rejction, on afamily of seven transmembrane G- rheumatoid arthritis or protein-coupledreceptors. Over 40 allergy human chemokines have been described, whichbind to ~17 receptors thus far identified. Chemokine GeneSeq WO9912968Chemokines are a family of related Chemokine activities can be Immunedisorders, hMCP1a Accession small, secreted proteins involved indetermined using assays known in Vascular disorders, Y14225 biologicalprocesses ranging from the art: Methods in Molecular Wound healing,cancer, hematopoiesis, agiogenesis, and Bilogy, 2000, vol. 138:Chemokine prevent organ transplant leukocye trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, rejection, Increase or familyare involved in a similarly T. N. C. Wells, and C. A. Power. enhance andiverse range of pathologies including © Humana Press Inc., inflammatoryresponse, inflammation, allergy, tissue rejection, Totowa, NJ viralkinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Chemokine GeneSeq WO9912968 Chemokinesare a family of related Chemokine activities can be Immune disorders,hMCP1b Accession small, secreted proteins involved in determined usingassays known in Vascular disorders, Y14226 biological processes rangingfrom the art: Methods in Molecular Wound healing, cancer, hematopoiesis,agiogenesis, and Bilogy, 2000, vol. 138: Chemokine prevent organtransplant leukocye trafficking: Members of this Protocols. Edited by:A. E. I. Proudfoot, rejection, Increase or family are involved in asimilarly T. N. C. Wells, and C. A. Power. enhance an diverse range ofpathologies including © Humana Press Inc., inflammatory response,inflammation, allergy, tissue rejection, Totowa, NJ viralk infection,and tumor biology. The chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Chemokine GeneSeq WO9912968 Chemokines are a family ofrelated Chemokine activities can be Immune disorders, hSDF1b Accessionsmall, secreted proteins involved in determined using assays known inVascular disorders, Y14228 biological processes ranging from the art:Methods in Molecular Wound healing, cancer, hematopoiesis, agiogenesis,and Bilogy, 2000, vol. 138: Chemokine prevent organ transplant leukocyetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,rejection, Increase or family are involved in a similarly T. N. C.Wells, and C. A. Power. enhance an diverse range of pathologiesincluding © Humana Press Inc., inflammatory response, inflammation,allergy, tissue rejection, Totowa, NJ viralk infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Chemokine GeneSeq WO9912968 Chemokines are a family ofrelated Chemokine activities can be Immune disorders, hIL-8 Accessionsmall, secreted proteins involved in determined using assays known inVascular disorders, Y14229 biological processes ranging from the art:Methods in Molecular Wound healing, cancer, hematopoiesis, agiogenesis,and Bilogy, 2000, vol. 138: Chemokine prevent organ transplant leukocyetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,rejection, Increase or family are involved in a similarly T. N. C.Wells, and C. A. Power. enhance an diverse range of pathologiesincluding © Humana Press Inc., inflammatory response, inflammation,allergy, tissue rejection, Totowa, NJ; and Holmes et al (1991) viralkinfection, and tumor biology. The Science 253, 1278-80. chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Chemokine GeneSeqWO9912968 Chemokines are a family of related Chemokine activities can beImmune disorders, hMCP1 Accession small, secreted proteins involved indetermined using assays known in Vascular disorders, Y14222 biologicalprocesses ranging from the art: Methods in Molecular Wound healing,cancer, hematopoiesis, agiogenesis, and Bilogy, 2000, vol. 138:Chemokine prevent organ transplant leukocye trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, rejection, Increase or familyare involved in a similarly T. N. C. Wells, and C. A. Power. enhance andiverse range of pathologies including © Humana Press Inc., inflammatoryresponse, inflammation, allergy, tissue rejection, Totowa, NJ viralkinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Chemokine GeneSeq WO9912968 Chemokinesare a family of related Chemokine activities can be Immune disorders,hMCP2 Accession small, secreted proteins involved in determined usingassays known in Vascular disorders, Y14223 biological processes rangingfrom the art: Methods in Molecular Wound healing, cancer, hematopoiesis,agiogenesis, and Bilogy, 2000, vol. 138: Chemokine prevent organtransplant leukocye trafficking. Members of this Protocols. Edited by:A. E. I. Proudfoot, rejection, Increase or family are involved in asimilarly T. N. C. Wells, and C. A. Power. enhance an diverse range ofpathologies including © Humana Press Inc., inflammatory response,inflammation, allergy, tissue rejection, Totowa, NJ viralk infection,and tumor biology. The chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Chemokine GeneSeq WO9912968 Chemokines are a family ofrelated Chemokine activities can be Immune disorders, hMCP3 Accessionsmall, secreted proteins involved in determined using assays known inVascular disorders, Y14224 biological processes ranging from the art:Methods in Molecular Wound healing, cancer, hematopoiesis, agiogenesis,and Bilogy, 2000, vol. 138: Chemokine prevent organ transplant leukocyetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,rejection, Increase or family are involved in a similarly T. N. C.Wells, and C. A. Power. enhance an diverse range of pathologiesincluding © Humana Press Inc., inflammatory response, inflammation,allergy, tissue rejection, Totowa, NJ viralk infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. C-C chemokine, GeneSeq EP905240 Chemokines are a family ofrelated Chemokine activities can be Inflammatory, Immune MCP2 Accessionsmall, secreted proteins involved in determined using assays known inand infectious diseases; Y05300 biological processes ranging from theart: Methods in Molecular pulmonary diseases and hematopoiesis,agiogenesis, and Bilogy, 2000, vol. 138: Chemokine skin disorders;tumours, leukocye trafficking. Members of this Protocols. Edited by: A.E. I. Proudfoot, and angiogenesis-and family are involved in a similarlyT. N. C. Wells, and C. A. Power. haematopoiesis-related diverse range ofpathologies including © Humana Press Inc., diseases inflammation,allergy, tissue rejection, Totowa, NJ viralk infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Wild type GeneSeq EP906954 Chemokines are a family ofrelated Chemokine activities can be Inflammatory, Immume monocyteAccession small, secreted proteins involved in determined using assaysknown in and infectious diseases; chemotactic Y07233 biologicalprocesses ranging from the art: Methods in Molecular pulmonary diseasesand protein 2 hematopoiesis, agiogenesis, and Bilogy, 2000, vol. 138:Chemokine skin disorders; tumours, leukocye trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, and angiogenesis-and familyare involved in a similarly T. N. C. Wells, and C. A. Power.haematopoiesis-related diverse range of pathologies including © HumanaPress Inc., diseases inflammation, allergy, tissue rejection, Totowa, NJviralk infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Truncated GeneSeq EP906954 Chemokinesarea family of related small, Chemokines activities can be Inflammatory,immune monocyte Accession secreted proteins involved in biologicaldetermined using assays known in the and infectious diseases;chemotactic Y07234 processes ranging from hematopoiesis, art: Methods inMolecular Biology, pulmonry diseases and protein 2 (6-76) angiogenesis,and leukocyte trafficking. 2000, vol. 138: Chemokine skin disorders;tumours, Members of this family are involved in a Protocols. Edited by:A. E. I. Proudfoot, and angiogenesis-and similarly diverse range ofpathologies T. N. C. Wells, and C. A. Power, haematopoiesis-relatedincluding inflammation, allergy, tissue Humana Press Inc., Totowa,diseases rejection, viral infection, and tumor NJ biology. Thechemokines exert their effects by acting on a fmaily of seventransmembrane G-protein-coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified.Truncated GeneSeq EP905241; Chemokines area family of related small,Chemokines activities can be Inflammatory, immune RANTES AccessionsEP906954 secreted proteins involved in biological determined usingassays known in the and infectious diseases; protein (3-68) Y07236 andprocesses ranging from hematopoiesis, art: Methods in Molecular Biology,pulmonry diseases and Y07232 angiogenesis, and leukocyte trafficking.2000, vol. 138: Chemokine skin disorders; tumours, Members of thisfamily are involved in a Protocols. Edited by: A. E. I. Proudfoot, andangiogenesis-and similarly diverse range of pathologies T. N. C. Wells,and C. A. Power, haematopoiesis-related including inflammation, allergy,tissue Humana Press Inc., Totowa, diseases rejection, viral infection,and tumor NJ biology. The chemokines exert their effects by acting on afmaily of seven transmembrane G-protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Wild type GeneSeq EP905241 Chemokines area family of relatedsmall, Chemokines activities can be Inflammatory, immune monocyteAccession secreted proteins involved in biological determined usingassays known in the and infectious diseases; chemotactic Y07237processes ranging from hematopoiesis, art: Methods in Molecular Biology,pulmonry diseases and protein 2 angiogenesis, and leukocyte trafficking.2000, vol. 138: Chemokine skin disorders; tumours, Members of thisfamily are involved in a Protocols. Edited by: A. E. I. Proudfoot, andangiogenesis-and similarly diverse range of pathologies T. N. C. Wells,and C. A. Power, haematopoiesis-related including inflammation, allergy,tissue Humana Press Inc., Totowa, diseases rejection, viral infection,and tumor NJ biology. The chemokines exert their effects by acting on afmaily of seven transmembrane G-protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Truncated GeneSeq EP905241 Chemokines area family of relatedsmall, Chemokines activities can be Inflammatory, immune monocyteAccession secreted proteins involved in biological determined usingassays known in the and infectious diseases; chemotactic Y07238processes ranging from hematopoiesis, art: Methods in Molecular Biology,pulmonry diseases and protein 2 (6-76) angiogenesis, and leukocytetrafficking. 2000, vol. 138: Chemokine skin disorders; tumours, Membersof this family are involved in a Protocols. Edited by: A. E. I.Proudfoot, and angiogenesis-and similarly diverse range of pathologiesT. N. C. Wells, and C. A. Power, haematopoiesis-related includinginflammation, allergy, tissue Humana Press Inc., Totowa, diseasesrejection, viral infection, and tumor NJ biology. The chemokines exerttheir effects by acting on a fmaily of seven transmembraneG-protein-coupled receptors. Over 40 human chemokines have beendescribed, which bind to ~17 receptors thus far identified. A partialGeneSeq EP897980 Chemokines area family of related small, Chemokinesactivities can be Soluble CXCR4B CXCR4B Accession secreted proteinsinvolved in biological determined using assays known in the receptorpolypeptides protein W97363 processes ranging from hematopoiesis, art:Methods in Molecular Biology, may be useful for angiogenesis, andleukocyte trafficking. 2000, vol. 138: Chemokine inhibiting chemokineMembers of this family are involved in a Protocols. Edited by: A. E. I.Proudfoot, activities and viral similarly diverse range of pathologiesT. N. C. Wells, and C. A. Power, infection. including inflammation,allergy, tissue Humana Press Inc., Totowa, rejection, viral infection,and tumor NJ biology. The chemokines exert their effects by acting on afamily of seven transmembrane G-protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Interferon GeneSeq U.S. Pat. No. 5,871,723 Chemokines areafamily of related small, Chemokines activities can be Angiogenesis,Cancer, gamma- Accession secreted proteins involved in biologicaldetermined using assays known in the Inflammatory and inducible W96709processes ranging from hematopoiesis, art: Methods in Molecular Biology,Immune disorders, protein (IP-10) angiogenesis, and leukocytetrafficking. 2000, vol. 138: Chemokine Cardio-Vascular Members of thisfamily are involved in a Protocols. Edited by: A. E. I. Proudfoot,discorders, Musco- similarly diverse range of pathologies T. N. C.Wells, and C. A. Power, skeletal disorders including inflammation,allergy, tissue Humana Press Inc., Totowa, rejection, viral infection,and tumor NJ biology. The chemokines exert their effects by acting on afmaily of seven transmembrane G-protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. A monokine GeneSeq U.S. Pat. No. 5,871,723 Chemokines areafamily of related small, Chemokines activities can be Angiogenesis,Cancer, induced by Accession secreted proteins involved in biologicaldetermined using assays known in the Inflammatory and gamma- W96710processes ranging from hematopoiesis, art: Methods in Molecular Biology,Immune disorders, interferon angiogenesis, and leukocyte trafficking.2000, vol. 138: Chemokine Cardio-Vascular (MIG) Members of this familyare involved in a Protocols. Edited by: A. E. I. Prougfoot; discorders,Musco- similarly diverse range of pathologies T. N. C. Wells, and C. A.Power, skeletal disorders including inflammation, allergy, tissue HumanaPress Inc., Totowa, rejection, viral infection, and tumor NJ biology.The chemokines exert their effects by acting on a fmaily of seventransmembrane G-protein-coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified.Interleukin-8 GeneSeq U.S. Pat. No. 5,871,723 Chemokines area family ofrelated small, Chemokines activities can be Angiogenesis, Cancer, (IL-8)protein. Accession secreted proteins involved in biological determinedusing assays known in the Inflammatory and W96711 processes ranging fromhematopoiesis, art: Methods in Molecular Biology, Immune disorders,angiogenesis, and leukocyte trafficking. 2000, vol. 138: ChemokineCardio-Vascular Members of this family are involved in a Protocols.Edited by: A. E. I. Proudfoot, discorders, Musco- similarly diverserange of pathologies T. N. C. Wells, and C. A. Power, skeletal disordersincluding inflammation, allergy, tissue Humana Press Inc., Totowa,rejection, viral infection, and tumor NJ; and Holmes et al (1991)Science biology. The chemokines exert their 253, 1278-80. effects byacting on a fmaily of seven transmembrane G-protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Epithelial GeneSeq U.S. Pat. No.5,871,723 Chemokines area family of related small, Chemokines activitiescan be Angiogenesis, Cancer, neutrophil Accession secreted proteinsinvolved in biological determined using assays known in the Inflammatoryand activating W96712 processes ranging from hematopoiesis, art: Methodsin Molecular Biology, Immune disorders, protein-78 angiogenesis, andleukocyte trafficking. 2000, vol. 138: Chemokine Cardio-Vascular(ENA-78) Members of this family are involved in a Protocols. Edited by:A. E. I. Proudfoot, discorders, Musco- similarly diverse range ofpathologies T. N. C. Wells, and C. A. Power, skeletal disordersincluding inflammation, allergy, tissue Humana Press Inc., Totowa,rejection, viral infection, and tumor NJ biology. The chemokines exerttheir effects by acting on a fmaily of seven transmembraneG-protein-coupled receptors. Over 40 human chemokines have beendescribed, which bind to ~17 receptors thus far identified. Growthrelated GeneSeq U.S. Pat. No. 5,871,723 Chemokines area family ofrelated small, Chemokines activities can be Angiogenesis, Cancer,oncogene-alpha Accession secreted proteins involved in biologicaldetermined using assays known in the Inflammatory and (GRO-alpha).W96713 processes ranging from hematopoiesis, art: Methods in MolecularBiology, Immune disorders, angiogenesis, and leukocyte trafficking.2000, vol. 138: Chemokine Cardio-Vascular Members of this family areinvolved in a Protocols. Edited by: A. E. I. Proudfoot, discorders,Musco- similarly diverse range of pathologies T. N. C. Wells, and C. A.Power, skeletal disorders including inflammation, allergy, tissue HumanaPress Inc., Totowa, rejection, viral infection, and tumor NJ biology.The chemokines exert their effects by acting on a fmaily of seventransmembrane G-protein-coupled receptors. Over 40 human chemokines havebeen described, which bind to ~17 receptors thus far identified. Growthrelated GeneSeq U.S. Pat. No. 5,871,723 Chemokines are a family ofrelated Chemokine activities can be Angiogenesis, Cancer, oncogene-betaAccession small, secreted proteins involved in determined using assaysknown in Inflammatory and (GRO-beta). W96714 biological processesranging from the art: Methods in Molecular Immune disorders,hematopoiesis, angiogenesis, and Biology, 2000, vol. 138: ChemokineCardio-Vascular leukocyte trafficking. Members of this Protocols. Editedby: A. E. I. Proudfoot, disorders, Musco- family are involved in asimilarly T. N. C. Wells, and C. A. Power, skeletal disorders diverserange of pathologies including © Humana Press Inc., inflammation,allergy, tissue rejection, Totowa, NJ viral infection and tumor biology.The chemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identifiedGrowth related GeneSeq U.S. Pat. No. 5,871,723 Chemokines are a familyof related Chemokine activities can be Angiogenesis, Cancer,oncogene-gamma Accession small, secreted proteins involved in determinedusing assays known in Inflammatory and (GRO-gamma) W96715 biologicalprocesses ranging from the art: Methods in Molecular Immune disorders,hematopoiesis, angiogenesis, and Biology, 2000, vol. 138: ChemokineCardio-Vascular leukocyte trafficking. Members of this Protocols. Editedby: A. E. I. Proudfoot, disorders, Musco- family are involved in asimilarly T. N. C. Wells, and C. A. Power, skeletal disorders diverserange of pathologies including © Humana Press Inc., inflammation,allergy, tissue rejection, Totowa, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. A platelet basic GeneSeq U.S. Pat. No. 5,871,723 Chemokinesare a family of related Chemokine activities can be Angiogenesis,Cancer, protein (PBP) Accession small, secreted proteins involved indetermined using assays known in Inflammatory and W96716 biologicalprocesses ranging from the art: Methods in Molecular Immune disorders,hematopoiesis, angiogenesis, and Biology, 2000, vol. 138: ChemokineCardio-Vascular leukocyte trafficking. Members of this Protocols. Editedby: A. E. I. Proudfoot, disorders, Musco- family are involved in asimilarly T. N. C. Wells, and C. A. Power, skeletal disorders diverserange of pathologies including © Humana Press Inc., inflammation,allergy, tissue rejection, Totowa, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Connective tissue GeneSeqAccession U.S. Pat. No. 5,871,723Chemokines are a family of related Chemokine activities can beAngiogenesis, Cancer, activating protein- S96717 small, secretedproteins involved in determined using assays known in Inflammatory andIII (CTAP-III) biological processes ranging from the art: Methods inMolecular Immune disorders, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine Cardio-Vascular leukocyte trafficking. Membersof this Protocols. Edited by: A. E. I. Proudfoot, disorders, Musco-family are involved in a similarly T. N. C. Wells, and C. A. Power,skeletal disorders diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Beta- GeneSeq U.S. Pat. No. 5,871,723Chemokines are a family of related Chemokine activities can beAngiogenesis, Cancer, thromboglobulin Accession small, secreted proteinsinvolved in determined using assays known in Inflammatory and protein(beta-TG) W96718 biological processes ranging from the art: Methods inMolecular Immune disorders, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine Cardio-Vascular leukocyte trafficking. Membersof this Protocols. Edited by: A. E. I. Proudfoot, disorders, Musco-family are involved in a similarly T. N. C. Wells, and C. A. Power,skeletal disorders diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Neutrophil GeneSeq U.S. Pat. No.5,871,723 Chemokines are a family of related Chemokine activities can beAngiogenesis, Cancer, activating peptide- Accession small, secretedproteins involved in determined using assays known in Inflammatory and 2(NAP-2) W96719 biological processes ranging from the art: Methods inMolecular Immune disorders, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine Cardio-Vascular leukocyte trafficking. Membersof this Protocols. Edited by: A. E. I. Proudfoot, disorders, Musco-family are involved in a similarly T. N. C. Wells, and C. A. Power,skeletal disorders diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Granulocyte GeneSeq U.S. Pat. No.5,871,723 Chemokines are a family of related Chemokine activities can beAngiogenesis, Cancer, chemotactic Accession small, secreted proteinsinvolved in determined using assays known in Inflammatory and protein-2(GCP-2) W96720 biological processes ranging from the art: Methods inMolecular Immune disorders, hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine Cardio-Vascular leukocyte trafficking. Membersof this Protocols. Edited by: A. E. I. Proudfoot, disorders, Musco-family are involved in a similarly T. N. C. Wells, and C. A. Power,skeletal disorders diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human chemokine GeneSeq EP887409Chemokines are a family of related Chemokine activities can be Immunedisorders, viral, MIG-beta protein Accession small, secreted proteinsinvolved in determined using assays known in parasitic, fungal or W90124biological processes ranging from the art: Methods in Molecularbacterial infections, hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine Cancer; autoimmune leukocyte trafficking. Members ofthis Protocols. Edited by: A. E. I. Proudfoot, diseases or transplantfamily are involved in a similarly T. N. C. Wells, and C. A. Power,rejection diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human ZCHEMO-8 GeneSeq WO9854326 Chemokines are a family ofrelated Chemokine activities can be Immune disorders, Accession small,secreted proteins involved in determined using assays known in cancer,myelopoietic W82716 biological processes ranging from the art: Methodsin Molecular disorders, autoimmune hematopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine disorders and leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot,immunodeficiencies, family are involved in a similarly T. N. C. Wells,and C. A. Power, Inflammatory and diverse range of pathologies including© Humana Press Inc., infectious diseases, inflammation, allergy, tissuerejection, Totowa, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human ZCHEMO-8 GeneSeq WO9854326 Chemokines are a familyof related Chemokine activities can be Immune disorders, Accessionsmall, secreted proteins involved in determined using assays known incancer, myelopoietic W82716 biological processes ranging from the art:Methods in Molecular disorders, autoimmune hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine disorders and leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,immunodeficiencies, family are involved in a similarly T. N. C. Wells,and C. A. Power, Inflammatory and diverse range of pathologies including© Humana Press Inc., infectious diseases, inflammation, allergy, tissuerejection, Totowa, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human Act-2 GeneSeq WO9854326 Chemokines are a family ofrelated Chemokine activities can be Immune disorders, protein Accessionsmall, secreted proteins involved in determined using assays known incancer, myelopoietic W82717 biological processes ranging from the art:Methods in Molecular disorders, autoimmune hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine disorders and leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,immunodeficiencies, family are involved in a similarly T. N. C. Wells,and C. A. Power, Inflammatory and diverse range of pathologies including© Humana Press Inc., infectious diseases, inflammation, allergy, tissuerejection, Totowa, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human SISD GeneSeq WO9854326 Chemokines are a family ofrelated Chemokine activities can be Immune disorders, protein Acessionsmall, secreted proteins involved in determined using assays known incancer, myelopoietic W82720 biological processes ranging from the art:Methods in Molecular disorders, autoimmune hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: disorders and leukocyte trafficking.Members of this Chemokine Protocols, Edited by: immunodeficiencies,family are involved in a similarly A. E. I. Proudfoot, T. N. C. Wells,and Inflammatory and diverse range of pathologies including C. A. Power.© Humana Press Inc., infectious diseases, inflammation, allergy, tissuerejection, Totowa, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human M110 GeneSeq WO9854326 Chemokines area family ofrelated Chemokine activities can be Immune disorders, protein Accessionsmall, secreted proteins involved in determined using assays known inthe cancer, myelopoietic W82721 biological processes ranging from art:Mehtods of Molecular Biology, disorders, autoimmune hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine disorders and leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,immunodeficiencies, family are involved in a similarly T. N. C. Wells,and C. A. Power Inflammatory and diverse range of pathologies including© Humana Press Inc., Totowa, infectious diseases, inflammation, allergy,tissue rejection, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human M11A GeneSeq WO9854326 Chemokines area family ofrelated Chemokine activities can be Immune disorders, protein Accessionsmall, secreted proteins involved in determined using assays known inthe cancer, myelopoietic W82722 biological processes ranging from art:Mehtods of Molecular Biology, disorders, autoimmune hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine disorders and leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,immunodeficiencies, family are involved in a similarly T. N. C. Wells,and C. A. Power Inflammatory and diverse range of pathologies including© Humana Press Inc., Totowa, infectious diseases, inflammation, allergy,tissue rejection, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. Human CCC3 GeneSeq WO9854326 Chemokines area family ofrelated Chemokine activities can be Immune disorders, protein Accessionsmall, secreted proteins involved in determined using assays known inthe cancer, myelopoietic W82723 biological processes ranging from art:Mehtods of Molecular Biology, disorders, autoimmune hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine disorders and leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,immunodeficiencies, family are involved in a similarly T. N. C. Wells,and C. A. Power Inflammatory and diverse range of pathologies including© Humana Press Inc., Totowa, infectious diseases, inflammation, allergy,tissue rejection, NJ Vascular disorders, viral infection, and tumorbiology. The wound healing chemokines exert their effects by acting on afamily of seven transmembrane G- protein-coupled receptors. Over 40human chemokines have been described, which bind to ~17 receptors thusfar identified. A human L105 GeneSeq WO9856818 Chemokines area family ofrelated Chemokine activities can be Cancer, wound healing chemokineAccession small, secreted proteins involved in determined using assaysknown in the designated W87588 biological processes ranging from art:Mehtods of Molecular Biology, huL105_3. hematopoiesis, angiogenesis, and2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power diverse range of pathologiesincluding © Humana Press Inc., Totowa, inflammation, allergy, tissuerejection, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. A human L105 GeneSeqWO9856818 Chemokines area family of related Chemokine activities can beCancer, wound healing chemokine Accession small, secreted proteinsinvolved in determined using assays known in the designated W87589biological processes ranging from art: Mehtods of Molecular Biology,huL105_7. hematopoiesis, angiogenesis, and 2000, vol. 138: Chemokineleukocyte trafficking. Members of this Protocols. Edited by: A. E. I.Proudfoot, family are involved in a similarly T. N. C. Wells, and C. A.Power diverse range of pathologies including © Humana Press Inc.,Totowa, inflammation, allergy, tissue rejection, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human mature GeneSeq WO9848828 Chemokines area family ofrelated Chemokine activities can be Infectious diseases, gro-alphaAccession small, secreted proteins involved in determined using assaysknown in the sepsis polypeptide W81498 biological processes ranging fromart: Mehtods of Molecular Biology, used to treat hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine sepsis leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Powerdiverse range of pathologies including © Humana Press Inc., Totowa,inflammation, allergy, tissue rejection, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human mature GeneSeq WO9848828 Chemokines area family ofrelated Chemokine activities can be Infectious diseases, gro-gammaAccession small, secreted proteins involved in determined using assaysknown in the sepsis polypeptide W81500 biological processes ranging fromart: Mehtods of Molecular Biology, used to treat hematopoiesis,angiogenesis, and 2000, vol. 138: Chemokine sepsis leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,family are involved in a similarly T. N. C. Wells, and C. A. Powerdiverse range of pathologies including © Humana Press Inc., Totowa,inflammation, allergy, tissue rejection, NJ viral infection, and tumorbiology. The chemokines exert their effects by acting on a family ofseven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human thymus GeneSeq WO0053635 Chemokines area family ofrelated Chemokine activities can be Inflammatory disorders, expressedAccessions small, secreted proteins involved in determined using assaysknown in the cancer, Immune and chemokine B19607 and biologicalprocesses ranging from art: Mehtods of Molecular Biology, vasculardisorders TECK and B19608 hematopoiesis, angiogenesis, and 2000, vol.138: Chemokine TECK variant leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power diverse range of pathologiesincluding © Humana Press Inc., Totowa, inflammation, allergy, tissuerejection, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human GeneSeq WO0042071Chemokines area family of related Chemokine activities can be Autoimmunedisorders, chemokine Accession B15791 small, secreted proteins involvedin determined using assays known in the Immune, Vascular and SDF1alphabiological processes ranging from art: Mehtods of Molecular Biology,Inflammatory disorders hematopoiesis, angiogenesis, and 2000, vol. 138:Chemokine leukocyte trafficking. Members of this Protocols. Edited by:A. E. I. Proudfoot, family are involved in a similarly T. N. C. Wells,and C. A. Power diverse range of pathologies including © Humana PressInc., Totowa, inflammation, allergy, tissue rejection, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, GROalpha Accession small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andB15793 biological processes ranging from the art: Methods in MolecularInflammatory diorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, eotaxin Accession small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andB15794 biological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, MIG Accession B15803 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, PF4 Accession B15804 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, I-309 Accession B15805 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, HCC-1 Accession B15806 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, C10 Accession B15807 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, CCR-2 Accession B15808 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, ENA-78 Accession B15809 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, GRObeta Accession B15810 small, secreted proteinsinvolved in determined using assasys known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot; family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, IP-10 Accession small, secreted proteins involvedin determined using assays known in Immune, Vascular and B15811biological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, SDF1beta Accession B15812 small, secreted proteinsinvolved in determined using assays known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, GRO alpha Accession B15813 small, secretedproteins involved in determined using assays known in Immune, Vascularand biological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. Human chemokine GeneSeq WO0042071Chemokines are a family of related Chemokine activities can beAutoimmune disorders, MIP1beta Accession B15831 small, secreted proteinsinvolved in determined using assays known in Immune, Vascular andbiological processes ranging from the art: Methods in MolecularInflammatory disorders hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine leukocyte trafficking. Members of this Protocols.Edited by: A. E. I. Proudfoot, family are involved in a similarly T. N.C. Wells, and C. A. Power. diverse range of pathologies including© Humana Press Inc., inflammation, allergy, tissue rejection, Totowa, NJviral infection, and tumor biology. The chemokines exert their effectsby acting on a family of seven transmembrane G- protein-coupledreceptors. Over 40 human chemokines have been described, which bind to~17 receptors thus far identified. A human C-C GeneSeq U.S. Pat. No.6,096,300 Chemokines are a family of related Chemokine activities can beCancer chemokine Accession B07939 small, secreted proteins involved indetermined using assays known in designated exodus biological processesranging from the art: Methods in Molecular hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine leukocyte trafficking. Members ofthis Protocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human chemokine GeneSeqU.S. Pat. No. 6,084,071 Chemokines are a family of related Chemokineactivities can be Chemotaxis, Gene L105_7 Accession Y96922 small,secreted proteins involved in determined using assays known in Therapy,Wound biological processes ranging from the art: Methods in Molecularhealing hematopoiesis, angiogenesis, and Biology, 2000, vol. 138:Chemokine leukocyte trafficking. Members of this Protocols. Edited by:A. E. I. Proudfoot, family are involved in a similarly T. N. C. Wells,and C. A. Power. diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human chemokine GeneSeq U.S. Pat. No.6,084,071 Chemokines are a family of related Chemokine activities can beChemotaxis, Gene L105_3 Accession Y96923 small, secreted proteinsinvolved in determined using assays known in Therapy, Wound biologicalprocesses ranging from the art: Methods in Molecular healinghematopoiesis, angiogenesis, and Biology, 2000, vol. 138: Chemokineleukocyte trafficking. Members of this Protocols. Edited by: A. E. I.Proudfoot, family are involved in a similarly T. N. C. Wells, and C. A.Power. diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified. Human secondary GeneSeq WO0038706 Chemokines are a family ofrelated Chemokine activities can be Cancer, Vascular and lymphoidAccession B01434 small, secreted proteins involved in determined usingassays known in Immune disorders chemokine (SLC) biological processesranging from the art: Methods in Molecular hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine leukocyte trafficking. Members ofthis Protocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human non-ELR GeneSeqWO0029439 Chemokines are a family of related Chemokine activities can beImmune and CXC chemokine Accession Y96310 small, secreted proteinsinvolved in determined using assays known in Inflammatory disorders,H174 biological processes ranging from the art: Methods in MolecularCancer, Haemostatic hematopoiesis, angiogenesis, and Biology, 2000, vol.138: Chemokine and thrombolytic leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, activity family are involvedin a similarly T. N. C. Wells, and C. A. Power. diverse range ofpathologies including © Humana Press Inc., inflammation, allergy, tissuerejection, Totowa, NJ viral infection, and tumor biology. The chemokinesexert their effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human non-ELR GeneSeqWO0029439 Chemokines are a family of related Chemokine activities can beImmune and CXC chemokine Accession Y96311 small, secreted proteinsinvolved in determined using assays known in Inflammatory disorders,IP10 biological processes ranging from the art: Methods in MolecularCancer, haemostatic and hematopoiesis, angiogenesis, and Biology, 2000,vol. 138: Chemokine thrombolytic activity leukocyte trafficking. Membersof this Protocols. Edited by: A. E. I. Proudfoot, family are involved ina similarly T. N. C. Wells, and C. A. Power. diverse range ofpathologies including © Humana Press Inc., inflammation, allergy, tissuerejection, Totowa, NJ viral infection, and tumor biology. The chemokinesexert their effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human non-ELR GeneSeqWO0029439 Chemokines are a family of related Chemokine activities can beImmune and CXC chemokine Accession small, secreted proteins involved indetermined using assays known in Inflammatory disorders, Mig Y96313biological processes ranging from the art: Methods in Molecular Cancer,haemostatic and hematopoiesis, angiogenesis, and Biology, 2000, vol.138: Chemokine thrombolytic activity leukocyte trafficking. Members ofthis Protocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human chemokine GeneSeqWO0028035 Chemokines are a family of related Chemokine activities can beCancer, wound healing, Ckbeta-7 Accession small, secreted proteinsinvolved in determined using assays known in inflammatory and Y96280biological processes ranging from the art: Methods in Molecularimmunoregulatory hematopoiesis, angiogenesis, and Biology, 2000, vol.138: Chemokine disorders leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human chemokine GeneSeqWO0028035 Chemokines are a family of related Chemokine activities can beCancer, wound healing, MIP-1alpha Accession small, secreted proteinsinvolved in determined using assays known in inflammatory and Y96281biological processes ranging from the art: Methods in Molecularimmunoregulatory hematopoiesis, angiogenesis, and Biology, 2000, vol.138: Chemokine disorders leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human mature GenSeqWO0028035 Chemokines are a family of related Chemokine activities can beCancer, wound healing, chemokine Accession small, secreted proteinsinvolved in determined using assays known in inflammatory and Ckbeta-7Y96282 biological processes ranging from the art: Methods in Molecularimmunoregulatory (optionally hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine disorders truncated) leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, family areinvolved in a similarly T. N. C. Wells, and C. A. Power. diverse rangeof pathologies including © Humana Press Inc., inflammation, allergy,tissue rejection, Totowa, NJ viral infection, and tumor biology. Thechemokines exert their effects by acting on a family of seventransmembrane G- protein-coupled receptors. Over 40 human chemokineshave been described, which bind to ~17 receptors thus far identified.Human chemokine GeneSeq WO0018431 Chemokines are a family of relatedChemokine activities can be Soluble CXCR3 receptor CXCR3 Accessionsmall, secreted proteins involved in determined using assays known inpolypeptides may be Y79372 biological processes ranging from the art:Methods in Molecular useful for inhibiting hematopoiesis, angiogenesis,and Biology, 2000, vol. 138: Chemokine chemokine activities leukocytetrafficking. Members of this Protocols. Edited by: A. E. I. Proudfoot,and viral infection. family are involved in a similarly T. N. C. Wells,and C. A. Power. diverse range of pathologies including © Humana PressInc., inflammation, allergy, tissue rejection, Totowa, NJ viralinfection, and tumor biology. The chemokines exert their effects byacting on a family of seven transmembrane G- protein-coupled receptors.Over 40 human chemokines have been described, which bind to ~17receptors thus far identified. Human neurotactin GeneSeq U.S. Pat. No.6,043,086 Chemokines are a family of related Chemokine activities can beNeurological disorders, chemokine like Accession small, secretedproteins involved in determined using assays known in Immune andrespiratory domain Y53259 biological processes ranging from the art:Methods in Molecular disorders hematopoiesis, angiogenesis, and Biology,2000, vol. 138: Chemokine leukocyte trafficking. Members of thisProtocols. Edited by: A. E. I. Proudfoot, family are involved in asimilarly T. N. C. Wells, and C. A. Power. diverse range of pathologiesincluding © Humana Press Inc., inflammation, allergy, tissue rejection,Totowa, NJ viral infection, and tumor biology. The chemokines exerttheir effects by acting on a family of seven transmembrane G-protein-coupled receptors. Over 40 human chemokines have been described,which bind to ~17 receptors thus far identified. Human CC type GeneSeqJP11302298 Chemokines are a family of related Chemokine activities canbe Cancer and infectious chemokine Accession small, secreted proteinsinvolved in determined using assays known in diseases interleukin CY57771 biological processes ranging from the art: Methods in Molecularhematopoiesis, angiogenesis, and Biology, 2000, vol. 138: Chemokineleukocyte trafficking. Members of this Protocols. Edited by: A. E. I.Proudfoot, family are involved in a similarly T. N. C. Wells, and C. A.Power. diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified Human CKbeta-9 GeneSeq U.S. Pat. No. 6,153,441 Chemokines area family of related Chemokine activities can be Cancer, Auto-immuneAccession small, secreted proteins involved in determined using assaysknown in and inflammatory B50860 biological processes ranging from theart: Methods in Molecular disorders, hematopoiesis, angiogenesis, andBiology, 2000, vol. 138: Chemokine Cardiovascular leukocyte trafficking.Members of this Protocols. Edited by: A. E. I. Proudfoot, disordersfamily are involved in a similarly T. N. C. Wells, and C. A. Power.diverse range of pathologies including © Humana Press Inc.,inflammation, allergy, tissue rejection, Totowa, NJ viral infection, andtumor biology. The chemokines exert their effects by acting on a familyof seven transmembrane G- protein-coupled receptors. Over 40 humanchemokines have been described, which bind to ~17 receptors thus faridentified Preproapolipoprotein GeneSeq WO9637608 Apoa-1 participates inthe reverse Lipid binding activity can be Useful for “paris” variantAccession transport of cholesterol from tissues to determined usingassays known in cardiovascular W08602 the liver for excretion bypromoting the art, such as, for example, the disorders, cholesterolcholesterol efflux from tissues and by Cholesterol Efflux Assays ofdisorders, and acting as a cofactor for the lecithin Takahaski et al.,P.N.A.S., Vol. 96, Hyperlipidaemia cholesterol acyltransferase (lcat).Issue 20, 11358-11363, Sep. 28, 1999. Preproapolipoprotein 5,721,114Apoa-1 participates in the reverse Lipid binding activity can be Usefulfor “milano” variant transport of cholesterol from tissues to determinedusing assays known in cardiovascular the liver for excretion bypromoting the art, such as, for example, the disorders, cholesterolcholesterol efflux from tissues and by Cholesterol Efflux Assays ofdisorders, and acting as a cofactor for the lecithin Takahaski et al.,P.N.A.S., Vol. 96, Hyperlipidaemia cholesterol acyltransferase (lcat).Issue 20, 11358-11363, Sep. 28, 1999. Glycodelin-A; GeneSeq WO9628169Naturally produced female contraceptive Glycodelin-A activity can beNaturally derived Progesterone- Accession that is removed rapidly fromthe body determined using the hemizona contraceptive useful forassociated W00289 following 2-3 days production. Uses assay as describedin Oehninger, S., the prevention of endometrial include contraceptionCoddington, C. C., Hodgen, G. D., pregnancy. protein and Seppala, M(1995) Fertil. Steril. 63, 377-383. NOGO-A Genbank NOGO polypeptides arepotent Inhibition of Neurite outgrowth. NOGO-A polypeptide Accessioninhibitors of neurite growth. Antagonists to NOGO polypeptidesantagonists are useful CAB99248 may promote the outgrowth of for thepromotion of neurites, thus inducing regeneration neural growth, whichof neurons. could be useful in the treatment of neural disorders anddysfunction due to degenerative diseases or trauma; useful in thetreatment of neoplastic diseases of the CNS; induce regeneration ofneurons or to promote the structural plasticity of the CNS. NOGO-BGenbank NOGO polypeptides are potent Inhibition of Neurite outgrowth.NOGO-B polypeptide Accession inhibitors of neurite growth. Antagoniststo NOGO polypeptides antagonists are useful CAB99249 may promote theoutgrowth of for the promotion of neurites, thus inducing regenerationneural growth, which of neurons. could be useful in the treatment ofneural disorders and dysfunction due to degenerative diseases or trauma;useful in the treatment of neoplastic diseases of the CNS; induceregeneration of neurons or to promote the structural plasticity of theCNS. NOGO-C Genbank NOGO polypeptides are potent Inhibition of Neuriteoutgrowth. NOGO-C polypeptide Accession inhibitors of neurite growth.Antagonists to NOGO polypeptides antagonists are useful CAB99250 maypromote the outgrowth of for the promotion of neurites, thus inducingregeneration neural growth, which of neurons. could be useful in thetreatment of neural disorders and dysfunction due to degenerativediseases or trauma; useful in the treatment of neoplastic diseases ofthe CNS; induce regeneration of neurons or to promote the structuralplasticity of the CNS. NOGO-66 Genbank NOGO polypeptides are potentInhibition of Neurite outgrowth by NOGO-66 receptor Receptor Accessioninhibitors of neurite growth, and are mediating the biological effectsof polypeptides are useful AAG53612 thought to mediate their effectsthrough NOGO polypeptides. Soluble for the promotion of the NOGO-66Receptor. NOGO-66 receptor polypeptides neural growth, which may promotethe outgrowth of could be useful in the neurites, thus inducingregeneration treatment of neural of neurons. disorders and dysfunctiondue to degenerative diseases or trauma; useful in the treatment ofneoplastic diseases of the CNS; induce regeneration of neurons or topromote the structural plasticity of the CNS. Antibodies specific U.S.Pat. No. 5,416,197 These antibodies are useful for the Collapsinactivity, which is thought Useful for the for collapsin promotion ofneurite outgrowth to inhibit the outgrowth of neurites, promotion ofneural can be assayed in the presence of growth, which could beantibodies specific for collapsing useful in the treatment using assaysknown in the art, such of neural disorders and as, for example, thecollapse assay dysfunction due to disclosed by Luo et al., Cell 1993degenerative diseases or Oct 22; 75(2): 217-27 trauma. Humanized Anti-WO9845331 These agents have anti-inflammatory VEGF activity can bedetermined Promotion of growth VEGF Antibodies, and anti-cancerapplications using assays known in the art, such and proliferation ofand fragments as those disclosed in International cells, such asvascular thereof Publication No. WO0045835, for endothelial cells.example. Antagonists may be useful as anti- angiogenic agents, and maybe applicable for cancer Humanized Anti- WO0029584 These agents haveanti-inflammatory VEGF activity can be determined Promotion of growthVEGF Antibodies, and anti-cancer applications using assays known in theart, such and proliferation of and fragments as those disclosed inInternational cells, such as vascular thereof Publication No. WO0045835,for endothelial cells. example. Antagonists may be useful as anti-angiogenic agents, and may be applicable for cancer Membrane boundGeneSeq. WO9963088 Cancer, Immune Disorders These proteins can be usedfor Activities can be proteins Accession linking bioactive molecules tocells determined using assay Y66631-Y66765 and for modulating biologicalknown in the art, activities of cells, using the suchas, for example,the polypeptides for specific targeting. assays disclosed in Thepolypeptide targeting can be International used to kill the targetcells, e.g. for Publication No. the treatment of cancers. TheseWO0121658. proteins are useful for the treatment of inunune systemdisorders. Secreted and GenSeq WO0053756 Cancer, Immune Disorders Theseproteins can be used for Activities can be Transmembrane Accessionlinking bioactive molecules to cells determined using assay polypeptidesB44241-B44334 and for modulating biological known in the art, activitiesof cells, using the suchas, for example, the polypeptides for specifictargeting. assays disclosed in The polypeptide targeting can beInternational used to kill the target cells, e.g. for Publication No.the treatment of cancers. These WO0121658 proteins are useful for thetreatment of immune system disorders. Secreted and GeneSeq WO9946281Cancer, Immune Disorders These proteins can be used for Activities canbe Transmembrane Accession linking bioactive molecules to cellsdetermined using assay polypeptides Y41685-Y41774 and for modulatingbiological known in the art, activities of cells, using the suchas, forexample, the polypeptides for specific targeting. assays disclosed inThe polypeptide targeting can be International used to kill the targetcells, e.g. for Publication No. the treatment of cancers. TheseWO0121658 proteins are useful for the treatment of immune systemdisorders.

β-Interferon

Most cytokines, including β-IFN, have relatively short circulationhalf-lives since they are produced in vivo to act locally andtransiently. To use β-IFN as an effective systemic therapeutic, oneneeds relatively large doses and frequent administrations. Such frequentparenteral administrations are inconvenient and painful. Further, toxicside effects are associated with β-IFN administration which are sosevere that some multiple sclerosis patients cannot tolerate thetreatment. These side effects are probably associated withadministration of a high dosage.

The present invention provides β-IFN/transferrin fusion proteins withincreased half-lives and pharmaceutical compositions comprising suchfusion proteins with increased stability. Such fusion proteins can beadministered to patients at lower doses, thus reducing the toxic sideeffects associated with β-IFN. The present invention contemplates theuse of the β-IFN/transferrin fusion proteins to treat various diseasesand conditions associated with β-IFN, such as but not limited tomultiple sclerosis, cancer including brain tumors and skin cancer, andviral infections such as hepatitis B and C. Preferably, theβ-IFN/transferrin fusion proteins are used to treat subjects sufferingfrom multiple sclerosis. As discussed below, formulations for oraladministration may also be produced.

β-interferon (β-IFN) is a glycoprotein with an apparent molecular weight(MW) of 23 kilodaltons. The gene encoding β-IFN is located on chromosome9. Its amino acid sequence containing 166 residues was determined by K.Hosoi et al. (J. Interferon Res., 8, pp 375-384 (1988)), and itsglucoside sequence was reported by Y. Kagawa et al. (J. Biol. Chem.,263, pp 17508-17515 (1988)).

β-IFN is secreted by fibroblasts in response to a viral or bacterialinfection, or exposure to foreign cells, macromolecules, or RNA. Inparticular, β-IFN inhibits the proliferation of infected cells andstimulates the immune system. The specific antiviral activity ofhomogeneous Hu-β-IFN is considered to be between 3×10⁸ and 1×10⁹ iu/mg(international units per milligram of total protein) inclusive (see U.S.Pat. No. 4,289,689 and EP-A-94 672).

“Interferon-beta” (IFN-β) or “beta-interferon” (β-IFN) includes nativeand recombinant Type I interferons exhibiting the same or similarpharmaceutical characteristics as the Type I interferons commonly knownas IFN-β-1a and IFN-β-1b.

Any β-IFN sequence may be used to prepare Tf fusion proteins of thepresent invention. For instance, U.S. Pat. No. 4,738,931 discloses thehuman β-IFN gene derived from human chromosomal DNA. A 1.8 kb EcoRIfragment, containing the nucleic acid encoding the human β-IFN,introduced into Escherichia coli has been deposited with the AmericanType Culture Collection in U.S.A. as Escherichia coli CI4 underaccession number ATCC 31905. The GenBank accession number for the aminoacid sequence of Human β-IFN amino acid sequence is AAA72588. The β-IFNcould also be a mutein as described in U.S. Pat. No. 4,588,585, in whichthe cysteine (Cys) normally occurring at position 17 of the wild-type ornative molecule has been replaced by a neutral amino acid, such asserine or alanine. Mark et al. (Proc. Natl. Acad. Sci. USA 81: 5662-5666(1984)) showed that when Cys 17 was changed for serine, the IFNexhibited the same spectrum of biological activities as β-IFN, such asanticellular and antiproliferative activity, activation of NK cells andneutralization of anti-human IFN antibodies. The mutein also exhibitedgreater stability than natural human (Hu) β-IFN when incubated at 70° C.

Because of its activity, β-IFN is regarded as an active principle notonly in the treatment and prophylaxis of viral diseases such as herpes,influenza etc, but also in the treatment of tumoral conditions such asencephaloma and leukemia. β-IFN is used to treat multiple sclerosis,brain tumor, skin cancer and hepatitis B and C. β-IFN fusion proteins ofthe present invention may be used to treat any of these diseases.

Human β-IFN is also effective in treating coronary restenosis in humansby selectively inhibiting the proliferation of coronary smooth musclecell at the site of vascular injury following a surgical procedure whilehaving no inhibitory effect on the normal proliferation of coronaryendothelial cells following the procedure. U.S. Pat. No. 5,681,558discloses a method of treating restenosis comprising administering β-IFNto the patient. Accordingly, β-IFN fusion proteins of the presentinvention may be used to treat restenosis.

β-IFN has an erythropoietic effect on the growth of progenitor cellsfrom individuals suffering from several diseases with a very lowproduction of red blood cells. Additionally, β-IFN increases burstformation as well as promotes a more rapid maturation toward normoblastsand even late reticulocytes. U.S. Pat. No. 5,104,653 discloses a methodfor the stimulation of erythropoiesis in a patient suffering from adisorder characterized by lack of maturation of progenitor blood cellsto red blood cells comprising administering to said patient anerythropoietic effective amount of human β-IFN. Therefore, β-IFN fusionproteins of the present invention may be used to stimulateerythropoiesis.

β-IFN, acting via STAT1 and STAT2, is known to upregulate anddownregulate a wide variety of genes, most of which are involved in theantiviral immune response. Although most IFN responses are induced bythe presence of dsRNA, both DNA and RNA viruses are sensitive to theeffects of β-IFN (Biron, Seminars in Immunology, 10: 383-390 (1998)).

β-IFN is generally produced in response to a viral infection. Interferonβ-IFN exerts its biological effects by binding to specific receptors onthe surface of human cells. This binding initiates a complex cascade ofintracellular events that leads to the expression of numerousinterferon-induced gene products and markers, for example,2′,5′-oligoadenylate synthetase, b₂-microglobulin, and neopterin.

(2′-5′)-Oligoadenylate synthetase and dsRNA dependent protein kinase arethe two best-known IFN-β-induced proteins (Biron, 1998, supra).(2′-5′)-oligoadenylate synthetase polymerizes ATP in a unique 2′-5′fashion (Janeway et al., Immunobiology: The Immune System in Health andDisease, 4th Edition, New York, Elsevier Science/Garland Publishing pp385-386 (1999)); the resultant oligomers activate RNase L, which cleavesmRNA (Biron, 1998, supra). dsRNA dependent protein kinase phosphorylatesand inactivates elF2, a transcriptional initiator. Both(2′-5′)-oligoadenylate synthetase and dsRNA dependent protein kinase actonly in the presence of dsRNA, i.e. in virally infected cells. The netresult of the action of these two proteins is to inhibit proteintranslation, which will retard viral replication (Biron, 1998, supra).

β-IFN dependent upregulation of TAP (transporter associated with antigenprocessing), Lmp2, Lmp7 serves to increase presentation of viralpeptides by MHC class I molecules in order to facilitate CD8 T cellrecognition and destruction of infected cells. TAP is the moleculeresponsible for loading peptide fragments onto MHC class I molecules inthe ER; the Lmp proteins are components of the proteasome which cleaveproteins specifically for MHC class I presentation (Janeway et al.,1999, supra).

β-IFN is known to both activate and induce some proliferation in naturalkiller (NK) cells (Janeway et al., 1999, supra). However, interferonsthemselves are not mitogens. The proliferation of NK cells is probablycaused by an intermediary cytokine which is induced by IFN-β (Biron,1998, supra). NK cells can kill cells which exhibit atypical patterns ofMHC class I expression; such cells are generally virally infected(Janeway et al., 1999, supra).

Although at the end of a successfully defeated infection, T cells die byapoptosis as the immune system returns to a homeostatic balance, some Tcells must avoid apoptosis and enter a G₀/G₁ memory state to preserveimmunological memory. These memory T cells are rescued from apoptosis byinteracting with stromal cells, which secrete β-IFN and some IFN-α(Pilling et al., European Journal of Immunology 29:1041-1050 (1999)). Tcell apoptosis may be induced by either cytokine deprivation or ligationof Fas on the cell surface, but β-IFN is able to block both apoptoticpathways. The former apoptotic pathway is blocked by β-IFN dependentupregulation of Bcl-x, an apoptotic inhibitor. Fas ligation-inducedapoptosis occurs much too quickly to be blocked by upregulation of agene, so β-IFN must block that apoptotic pathway by different means(Scheel-Toellner et al., European Journal of Immunology 29:2603-2612(1999)). The existence of a second blocking mechanism is supported bythe results of Marrack et al. (Journal of Experimental Medicine189:521-529 (1999)), who found that β-IFN prevented T cell apoptosiswithout increased production of Bcl-x.

Der et al. (Proc. Nat. Acad. Sci. USA 95: 15623-15628 (1998)) found thatβ-IFN increased transcription of well over 100 proteins in humanfibrosarcoma cells. Induced proteins ranged in function from cytochromesand cell scaffolding proteins to immunologically active proteins such asComplement components and dsRNA adenosine deaminase. These resultsindicate that β-IFN has truly pleiotropic effects, Many of which are notfully understood.

Much clinical research on β-IFN is currently focused on its use as atreatment for multiple sclerosis (MS). MS is an autoimmune disease inwhich T cells mount an immune response against self myelin antigens inthe glial cells of the central nervous system (Goodkin, 1999. Multiplesclerosis: Treatment options for patients with relapsing-remitting andsecondary progressive multiple sclerosis.<http://www.msnews.org/goodkin1_(—)99.htm>). In 1993, the FDA approvedsubcutaneous injections of IFN-β1b for treatment of MS (Revelle M.,1993, FDA licenses interferon beta-1b.(<http://www.fda.gov/bbs/topics/NEWS/NEW00424.html>). β-IFN 1b is anon-glycosylated form of IFN-β produced in E. coli (Arduini et al.,Protein Science 8: 1867-1877 (1999)). Adverse experiences associatedwith β-IFN 1b therapy include: injection site reactions (inflammation,pain, hypersensitivity and necrosis), and a flu-like symptom complex(fever, chills, anxiety and confusion). These adverse side effects maybe, in fact, reduced or alleviated by fusing β-IFN 1b to transferrin asdescribed above.

Currently, β-IFN 1a (an eukaryotic, glycosylated form) is also available(Goodkin, 1999, supra). β-IFN 1a is produced by recombinant DNAtechnology. Interferon beta-1a is a 166 amino acid glycoprotein with apredicted molecular weight of approximately 22,500 daltons. It isproduced by mammalian cells (Chinese Hamster Ovary cells) into which thehuman IFN-β gene has been introduced. The amino acid sequence of β-IFN1a is identical to that of natural human β-IFN and may be used to makeTf fusion proteins of the present invention.

β-IFN/transferrin fusion proteins treatment may also ameliorateautoimmune attacks by restoring suppressor T cell function; cotreatmentwith all-trans-retinoic acid seems to increase this restorative actionfor unknown reasons (Qu et al., 1998. All-trans retinoic acidpotentiates the ability of interferon beta-1b.<http://members.tripod.com/˜ThJuland/ra-beta1b.html>). β-IFN may alsoinhibit the induction of inducible nitric oxide synthase (INOS)expression by IL-1 and IFN-γ. Production of nitric oxide by INOS inastrocytes has been implicated as a factor in the parthenogenesis of MS(Hua et al. 1998. Beta inteferon prevents nitric oxide/peroxynitratefrom damaging the central nervous system.(<http://members.tripod.com/˜ThJuland/nitric-oxide_beta.html>).

In one aspect, the present invention includes the use of β-IFN analogsthat are therapeutically effective for treating various diseasesassociated with β-IFN for generating β-IFN/transferrin fusion proteins.

In another aspect, the present invention includes the use of theβ-IFN/transferrin fusion protein in the methods described above toinhibit or stimulate various cellular processes and for the treatmentand prevention of the various disease and conditions described above. Inparticular, the β-IFN/transferrin fusion protein may be used to treatmultiple sclerosis, herpes, influenza, brain tumor, and skin cancer.

The β-IFN/transferrin fusion protein of the present invention can beformulated into pharmaceutical compositions by well known methods. See,e.g., Remington's Pharmaceutical Sciences by E. W. Martin, herebyincorporated by reference, describes suitable formulations. Thepharmaceutical composition of the β-IFN/transferrin fusion protein ofthe present invention may be formulated in a variety of forms, includingliquid, gel, lyophilized, or any other suitable form. The preferred formwill depend upon the particular indication being treated and will beapparent to one of skill in the art

The β-IFN/transferrin fusion protein can be administered in pure form orin an appropriate pharmaceutical composition. Administration can becarried out via any of the accepted modes. Thus, administration can be,for example, orally, nasally, parenterally, topically, transdermally, orrectally, in the form of solid, semi-solid, lyophilized powder, orliquid dosage forms, such as for example, tablets, suppositories, pills,soft elastic and hard gelatin capsules, powders, solutions, suspensions,or aerosols, or the like, preferably in unit dosage forms suitable forsimple administration of precise dosages. The compositions will includea conventional pharmaceutical carrier or excipient and theβ-IFN/transferrin fusion protein as the active agent, and, in addition,may include other medicinal agents, pharmaceutical agents, carriers,adjuvants, etc.

Generally, depending on the intended mode of administration, thepharmaceutically acceptable compositions will contain about 1% to about99% by weight of the β-IFN/transferrin fusion protein, and 99% to 1% byweight of a suitable pharmaceutical excipient. The composition could beabout 5% to 75% by weight of the β-IFN/transferrin fusion protein withthe rest being suitable pharmaceutical excipients.

The route of administration could be parenterally, using a convenientdaily dosage regimen which can be adjusted according to the degree ofseverity of the disease, preferably multiple sclerosis, to be treated.For such parenteral administration, a pharmaceutically acceptablecomposition containing the β-IFN/transferrin fusion protein may beformed by the methods disclosed in U.S. Pat. Nos. 4,462,940, 4,588,585and 4,992,271.

Alternatively, the β-IFN/transferrin fusion protein pharmaceuticalcompositions may be administered orally, intravenously, intramuscularly,intraperitoneally, intradermally or subcutaneously or in any otheracceptable manner. The preferred mode of administration will depend uponthe particular indication being treated and will be apparent to one ofskill in the art.

U.S. Pat. No. 6,333,032 describes effective methods of using β-IFN totreat diseases in warm-blooded vertebrates, such as multiple sclerosis.Treatment of multiple sclerosis comprises administering β-IFN at adosage of 0.01 to about 5 IU/lb per day in a dosage form adapted topromote contact of said dosage of interferon with the oral andpharyngeal mucosa of said animal. The dosage of interferon could be from0.1 to about 4.0 IU/lb per day, or from 0.5 to about 1.5 IU/lb of bodyweight per day.

The present invention contemplates administering the β-IFN in a dosageform adapted to assure maximum contact of the interferon in said dosageform with the oral and pharyngeal mucosa of the human or animalundergoing treatment. Contact of interferon with the mucosa can beenhanced by maximizing residence time of the treatment solution in theoral or pharyngeal cavity. Thus, best results seem to be achieved inhuman patients when the patient is requested to hold said solution ofinterferon in the mouth for a period of time. Contact of interferon withthe oral and pharyngeal mucosa and thereafter with the lymphatic systemof the treated human or animal is unquestionably the most efficientmethod administering immunotherapeutic amounts of interferon.

Further, the present invention contemplates the use of theβ-IFN/transferrin protein for the manufacture of a medicament which isuseful for the treatment of diseases associated with β-IFN. The diseasescontemplated by the present invention include but are not limited tothose described above.

Glucagon-Like Peptide-1 (GLP-1)

Glucagon-Like Peptide-1 (GLP-1) is a gastrointestinal hormone thatregulates insulin secretion belonging to the so-called enteroinsularaxis. The enteroinsular axis designates a group of hormones, releasedfrom the gastrointestinal mucosa in response to the presence andabsorption of nutrients in the gut, which promote an early andpotentiated release of insulin. The incretin effect which is theenhancing effect on insulin secretion is probably essential for a normalglucose tolerance. GLP-1 is a physiologically important insulinotropichormone because it is responsible for the incretin effect.

GLP-1 is a product of proglucagon (Bell, et al., Nature, 1983, 304:368-371). It is synthesized in intestinal endocrine cells in twoprincipal major molecular forms, as GLP-1(7-36)amide and GLP-1(7-37).The peptide was first identified following the cloning of cDNAs andgenes for proglucagon in the early 1980s.

Initial studies done on the full length peptides GLP-1(1-37) andGLP-1(1-36^(amide)) concluded that the larger GLP-1 molecules are devoidof biological activity. In 1987, three independent research groupsdemonstrated that removal of the first six amino acids resulted in aGLP-1 molecule with enhanced biological activity.

The amino acid sequence of GLP-1 is disclosed by Schmidt et al. (1985Diabetologia 28 704-707). Human GLP-1 is a 37 amino acid residue peptideoriginating from preproglucagon which is synthesized in the L-cells inthe distal ileum, in the pancreas, and in the brain. Processing ofpreproglucagon to GLP-1(7-36^(amide)), GLP-1(7-37) and GLP-2 occursmainly in the L-cells. The amino acid sequence of GLP-1(7-36^(amide))and GLP-1(7-37) is (SEQ ID NO: 6):

His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Xwherein X is NH₂ for GLP-1(7-36^(amide)) and X is Gly for GLP-1(7-37).

GLP-1 like molecules possesses anti-diabetic activity in human subjectssuffering from Type II (non-insulin-dependent diabetes mellitus (NIDDM))and, in some cases, even Type I diabetes. Treatment with GLP-1 elicitsactivity, such as increased insulin secretion and biosynthesis, reducedglucagon secretion, delayed gastric emptying, only at elevated glucoselevels, and thus provides a potentially much safer therapy than insulinor sulfonylureas. Post-prandial and glucose levels in patients can bemoved toward normal levels with proper GLP-1 therapy. There are alsoreports suggesting GLP-1-like molecules possess the ability to preserveand even restore pancreatic beta cell function in Type-II patients.

Any GLP-1 sequence may be used to make Tf fusion proteins of the presentinvention, including GLP-1(7-35), GLP-1(7-36), and GLP-1(7-37). GLP-1also has powerful actions on the gastrointestinal tract. Infused inphysiological amounts, GLP-1 potently inhibits pentagastrin-induced aswell as meal-induced gastric acid secretion (Schjoldager et al., Dig.Dis. Sci. 1989, 35:703-708; Wettergren et al., Dig Dis Sci 1993;38:665-673). It also inhibits gastric emptying rate and pancreaticenzyme secretion (Wettergren et al., Dig Dis Sci 1993; 38:665-673).Similar inhibitory effects on gastric and pancreatic secretion andmotility may be elicited in humans upon perfusion of the ileum withcarbohydrate- or lipid-containing solutions (Layer et al., Dig Dis Sci1995, 40:1074-1082; Layer et al., Digestion 1993, 54: 385-38).Concomitantly, GLP-1 secretion is greatly stimulated, and it has beenspeculated that GLP-1 may be at least partly responsible for thisso-called “ileal-brake” effect (Layer et al., Digestion 1993; 54:385-38). In fact, recent studies suggest that, physiologically, theileal-brake effects of GLP-1 may be more important than its effects onthe pancreatic islets. Thus, in dose response studies GLP-1 influencesgastric emptying rate at infusion rates at least as low as thoserequired to influence islet secretion (Nauck et al., Gut 1995; 37(suppl. 2): A124).

GLP-1 seems to have an effect on food intake. Intraventricularadministration of GLP-1 profoundly inhibits food intake in rats (Schicket al. in Ditschuneit et al. (eds.), Obesity in Europe, John Libbey &Company ltd, 1994; pp. 363-367; Turton et al., Nature 1996, 379: 69-72).This effect seems to be highly specific. Thus, N-terminally extendedGLP-1(PG 72-107) amide is inactive and appropriate doses of the GLP-1antagonist, exendin 9-39, abolish the effects of GLP-1(Tang-Christensenet al., Am. J. Physiol., 1996, 271(4 Pt 2):R848-56). Acute, peripheraladministration of GLP-1 does not inhibit food intake acutely in rats(Tang-Christensen et al., Am. J. Physiol., 1996, 271(4 Pt 2):R848-56;Turton et al., Nature 1996, 379: 69-72). However, it remains possiblethat GLP-1 secreted from the intestinal L-cells may also act as asatiety signal.

In diabetic patients, GLP's insulinotropic effects and the effects ofGLP-1 on the gastrointestinal tract are preserved (Willms et al,Diabetologia 1994; 37, suppl.1: A118), which may help curtailmeal-induced glucose excursions, but, more importantly, may alsoinfluence food intake. Administered intravenously, continuously for oneweek, GLP-1 at 4 ng/kg/min has been demonstrated to dramatically improveglycaemic control in NIDDM patients without significant side effects(Larsen et al., Diabetes 1996; 45, suppl. 2: 233A.).

GLP-1/transferrin fusion proteins comprising at least one analog ofGLP-1 and fragments thereof are useful in the treatment of Type 1 andType 2 diabetes and obesity and may be formulated for oraladministration as described below.

As used herein, the term “GLP-1 molecule” means GLP-1, a GLP-1 analog,or GLP-1 derivative.

As used herein, the term “GLP-1 analog” is defined as a molecule havingone or more amino acid substitutions, deletions, inversions, oradditions compared with GLP-1. Many GLP-1 analogs are known in the artand include, for example, GLP-1(7-34), GLP-1(7-35), GLP-1(7-36),Val⁸-GLP-1(7-37), Gly⁸-GLP-1(7-37), Ser⁸-GLP-1(7-37), Gln⁹-GLP1(7-37),D-Gln⁹-GLP-1(7-37), Thr¹⁶-Lys¹⁸-GLP-1(7-37), and Lys¹⁸-GLP-1(7-37).Other analogs include dipeptidyl-peptidase resistant versions of GLP-1,wherein the N-terminal end of the peptide is protected. Such analogsinclude, but are not limited to GLP-1 with additional amino acids, suchas histidine residue added to the N-terminal end or substituted into theN-terminal amino acids (amino acid 7 or 8 in GLP-1(7-36) or GLP-1(7-37).In these analogs, the N-terminal end may comprise the residuesHis-His-Ala, Gly-His-Ala, His-Gly-Glu, His-Ser-Glu, His-Ala-Glu,His-Gly-Glu, His-Ser-Glu, His-His-Ala-Glu, His-His-Gly-Glu,His-His-Ser-Glu, Gly-His-Ala-Glu, Gly-His-Gly-Glu, Gly-His-Ser-Glu,His-X-Ala-Glu, His-X-Gly-Glu, His-X-Ser-Glu, wherein X is any aminoacid. U.S. Pat. No. 5,118,666 discloses examples of GLP-1 analogs suchas GLP-1(7-34) and GLP-1(7-35).

The term “GLP-1 derivative” is defined as a molecule having the aminoacid sequence of GLP-1 or a GLP-1 analog, but additionally havingchemical modification of one or more of its amino acid side groups,α-carbon atoms, terminal amino group, or terminal carboxylic acid group.A chemical modification includes, but is not limited to, adding chemicalmoieties, creating new bonds, and removing chemical moieties.

As used herein, the term “GLP-1 related compound” refers to any compoundfalling within the GLP-1, GLP-1 analog, or GLP-1 derivative definition.

WO 91/11457 discloses analogs of the active GLP-1 peptides 7-34, 7-35,7-36, and 7-37 which can also be useful as GLP-1 moieties.

EP 0708179-A2 (Eli Lilly & Co.) discloses GLP-1 analogs and derivativesthat include an N-terminal imidazole group and optionally an unbranchedC₆-C₁₀ acyl group in attached to the lysine residue in position 34.

EP 0699686-A2 (Eli Lilly & Co.) discloses certain N-terminal truncatedfragments of GLP-1 that are reported to be biologically active.

U.S. Pat. No. 5,545,618 discloses GLP-1 molecules consisting essentiallyof GLP-1(7-34), GLP1(7-35), GLP-1(7-36), or GLP-1(7-37), or the amideforms thereof, and pharmaceutically-acceptable salts thereof, having atleast one modification selected from the group consisting of: (a)substitution of glycine, serine, cysteine, threonine, asparagine,glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine,phenylalanine, arginine, or D-lysine for lysine at position 26 and/orposition 34; or substitution of glycine, serine, cysteine, threonine,asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine,methionine, phenylalanine, lysine, or a D-arginine for arginine atposition 36; (b) substitution of an oxidation-resistant amino acid fortryptophan at position 31; (c) substitution of at least one of: tyrosinefor valine at position 16; lysine for serine at position 18; asparticacid for glutamic acid at position 21; serine for glycine at position22; arginine for glutamine at position 23; arginine for alanine atposition 24; and glutamine for lysine at position 26; and (d)substitution of at least one of: glycine, serine, or cysteine foralanine at position 8; aspartic acid, glycine, serine, cysteine,threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine,leucine, methionine, or phenylalanine for glutamic acid at position 9;serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine,valine, isoleucine, leucine, methionine, or phenylalanine for glycine atposition 10; and glutamic acid for aspartic acid at position 15; and (e)substitution of glycine, serine, cysteine, threonine, asparagine,glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine,or phenylalanine, or the D- or N-acylated or alkylated form of histidinefor histidine at position 7; wherein, in the substitutions is (a), (b),(d), and (e), the substituted amino acids can optionally be in theD-form and the amino acids substituted at position 7 can optionally bein the N-acylated or N-alkylated form.

U.S. Pat. No. 5,118,666 discloses a GLP-1 molecule having insulinotropicactivity. Such molecule is selected from the group consisting of apeptide having the amino acid sequenceHis-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys(SEQ ID NO: 7) orHis-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly(SEQ ID NO: 8); and a derivative of said peptide and wherein saidpeptide is selected from the group consisting of: apharmaceutically-acceptable acid addition salt of said peptide; apharmaceutically-acceptable carboxylate salt of said peptide; apharmaceutically-acceptable lower alkylester of said peptide; and apharmaceutically-acceptable amide of said peptide selected from thegroup consisting of amide, lower alkyl amide, and lower dialkyl amide.

U.S. Pat. No. 6,277,819 teaches a method of reducing mortality andmorbidity after myocardial infarction comprising administering GLP-1,GLP-1 analogs, and GLP-1 derivatives to the patient. The GLP-1 analogbeing represented by the following structural formula (SEQ ID NO: 9):R₁-X₁-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-X₂-Gly-Gln-Ala-Ala-Lys-X₃-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-R₂and pharmaceutically-acceptable salts thereof, wherein: R₁ is selectedfrom the group consisting of L-histidine, D-histidine,desamino-histidine, 2-amino-histidine, .beta.-hydroxy-histidine,homohistidine, alpha-fluoromethyl-histidine, and alpha-methyl-histidine;X₁ is selected from the group consisting of Ala, Gly, Val, Thr, Ile, andalpha-methyl-Ala; X₂ is selected from the group consisting of Glu, Gln,Ala, Thr, Ser, and Gly; X₃ is selected from the group consisting of Glu,Gln, Ala, Thr, Ser, and Gly; R₂ is selected from the group consisting ofNH₂, and Gly-OH; provided that the GLP-1 analog has an isoelectric pointin the range from about 6.0 to about 9.0 and further providing that whenR₁ is His, X₁ is Ala, X₂ is Glu, and X₃ is Glu, must be NH₂.

Ritzel et al. (Journal of Endocrinology, 1998, 159: 93-102) disclose aGLP-1 analog, [Ser⁸]GLP-1, in which the N-terminal second amino acid,alanine, is replaced with serine. The modification did not impair theinsulinotropic action of the peptide but produced an analog withincreased plasma stability as compared to GLP-1.

U.S. Pat. No. 6,429,197 teaches that GLP-1 treatment after acute strokeor hemorrhage, preferably intravenous administration, can be an idealtreatment because it provides a means for optimizing insulin secretion,increasing brain anabolism, enhancing insulin effectiveness bysuppressing glucagon, and maintaining euglycemia or mild hypoglycemiawith no risk of severe hypoglycemia or other adverse side effects. Thepresent invention provides a method of treating the ischemic orreperfused brain with GLP-1 or its biologically active analogues afteracute stroke or hemorrhage to optimize insulin secretion, to enhanceinsulin effectiveness by suppressing glucagon antagonism, and tomaintain euglycemia or mild hypoglycemia with no risk of severehypoglycemia.

U.S. Pat. No. 6,277,819 provides a method of reducing mortality andmorbidity after myocardial infarction, comprising administering to apatient in need thereof, a compound selected from the group consistingof GLP-1, GLP-1 analogs, GLP-1 derivatives andpharmaceutically-acceptable salts thereof, at a dose effective tonormalize blood glucose.

U.S. Pat. No. 6,191,102 discloses a method of reducing body weight in asubject in need of body weight reduction by administering to the subjecta composition comprising a glucagon-like peptide-1 (GLP-1), aglucagon-like peptide analog (GLP-1 analog), a glucagon-like peptidederivative (GLP-1 derivative) or a pharmaceutically acceptable saltthereof in a dose sufficient to cause reduction in body weight for aperiod of time effective to produce weight loss, said time being atleast 4 weeks.

GLP-1 is fully active after subcutaneous administration (Ritzel et al.,Diabetologia 1995; 38: 720-725), but is rapidly degraded mainly due todegradation by dipeptidyl peptidase IV-like enzymes (Deacon et al., JClin Endocrinol Metab 1995, 80: 952-957; Deacon et al., 1995, Diabetes44: 1126-1131). Thus, unfortunately, GLP-1 and many of its analogueshave a short plasma half-life in humans (Orskov et al., Diabetes 1993;42:658-661). Accordingly, it is an objective of the present invention toprovide transferrin fusion proteins comprising GLP-1 or analoguesthereof which have a protracted profile of action relative toGLP-1(7-37). It is a further object of the invention to providederivatives of GLP-1 and analogues thereof which have a lower clearancethan GLP-1(7-37). Moreover, it is an object of the invention to providepharmaceutical compositions comprising GLP-1/transferrin fusion proteinsor GLP-1 analog/transferrin fusion proteins with improved stability.Additionally, the present invention includes the use ofGLP-1/transferrin fusion proteins or GLP-1 analog/transferrin fusionproteins to treat diseases associated with GLP-1 such as but not limitedto those described above.

In one aspect of the present invention, the pharmaceutical compositionscomprising the GLP-1 peptide/transferrin fusion proteins and GLP-1analog/transferrin fusion proteins may be formulated by any of theestablished methods of formulating pharmaceutical compositions, e.g. asdescribed in Remington's Pharmaceutical Sciences, 1985. The compositionmay be in a form suited for systemic injection or infusion and may, assuch, be formulated with a suitable liquid vehicle such as sterile wateror an isotonic saline or glucose solution. The compositions may besterilized by conventional sterilization techniques which are well knownin the art. The resulting aqueous solutions may be packaged for use orfiltered under aseptic conditions and lyophilized, the lyophilizedpreparation being combined with the sterile aqueous solution prior toadministration. The composition may contain pharmaceutically acceptableauxiliary substances as required to approximate physiologicalconditions, such as buffering agents, tonicity adjusting agents and thelike, for instance sodium acetate, sodium lactate, sodium chloride,potassium chloride, calcium chloride, etc.

The GLP-1/transferrin fusion proteins and GLP-1 analog/transferrinfusion proteins of the present invention may also be adapted for oral,nasal, transdermal, pulmonal or rectal administration. Thepharmaceutically acceptable carrier or diluent employed in thecomposition may be any conventional solid carrier. Examples of solidcarriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin,acacia, magnesium stearate and stearic acid. Similarly, the carrier ordiluent may include any sustained release material known in the art,such as glyceryl monostearate or glyceryl distearate, alone or mixedwith a wax.

It may be of particular advantage to provide the composition of theinvention in the form of a sustained release formulation. As such, thecomposition may be formulated as microcapsules or microparticlescontaining the GLP-1/transferrin fusion protein or GLP-1analog/transferrin fusion protein encapsulated by or dispersed in asuitable pharmaceutically acceptable biodegradable polymer such aspolylactic acid, polyglycolic acid or a lactic acid/glycolic acidcopolymer.

For nasal administration, the preparation may contain GLP-1/transferrinfusion proteins or GLP-1 analog/transferrin fusion proteins dissolved orsuspended in a liquid carrier, in particular an aqueous carrier, foraerosol application. The carrier may contain additives such assolubilizing agents, e.g. propylene glycol, surfactants, absorptionenhancers such as lecithin (phosphatidylcholine) or cyclodextrin, orpreservatives such as parabenes.

Generally, the compounds of the present invention are dispensed in unitdosage form comprising 0.5-500 mg of the fusion protein together with apharmaceutically acceptable carrier per unit dosage.

Moreover, the present invention contemplates the use of theGLP-1/transferrin and GLP-1 analog/transferrin fusion proteins for themanufacture of a medicinal product which can be used in the treatment ofdiseases associated with elevated glucose level, such as but not tolimited to those described above. Specifically, the present inventioncontemplates the use of GLP-1/transferrin fusion protein for thetreatment of diabetes including type II diabetes, obesity, severe burns,and heart failure, including congestive heart failure and acute coronarysyndrome.

The N-terminus of GLP-1 is normally amidated. In yeast, amidation doesnot occur. In one aspect of the invention, in order to compensate foramidation on the N-terminus which does not occur in yeast, an extraamino acid is added on the N-terminus of GLP-1. The addition of an aminoacid to the N-terminus of GLP-1 may prevent dipeptidyl peptidase fromcleaving at the second amino acid of GLP-1 due to steric hindrance.Therefore, GLP-1 will remain functionally active. Any one of the 20amino acids may be added to the N-terminus of GLP-1. In some instances,an uncharged or positively charged amino acid may be used andpreferably, a smaller amino acid such as Glycine is added. The GLP-1with the extra amino acid is then fused to transferrin. Accordingly, theGLP-1 with the added amino acid will be fused at the N-terminus of theGLP-1/transferrin fusion protein.

In one embodiment of making the GLP-1(7-36) or GLP-1(7-37) peptide moreresistant to dipeptidyl peptidase, a His residue is added at theN-terminus of GLP-1 or is inserted after the His residue at theN-terminus of GLP-1, so that the N-terminus of GLP-1 begins withHis-His.

In another embodiment of the invention, the second residue from theN-terminus in the GLP-1(7-36) or GLP-1(7-37) peptide (SEQ ID NO: 6) issubstituted with another amino acid. For example, the Ala residue at thesecond residue from the N-terminus in the GLP-1(7-36) or GLP-1(7-37)peptide may be substituted with Ser, Gly, Val, or another amino acid.

GLP-mTf Fusion Protein for Treating Type 2 Diabetes

As discussed above, GLP-1 activates and regulates important endocrinehormone systems in the body and plays a critical management role in themetabolism of glucose. Unlike all other diabetic treatments on themarket GLP-1 has the potential to be restorative by acting as a growthfactor for B-cells thus improving the ability of the pancreas to secreteinsulin and also, to make the existing insulin levels act moreefficiently by improving sensitivity and better stabilizing glucoselevels. This reduces the burden on daily monitoring of glucose levelsand potentially offers a delay in the serious long term side effectscaused by fluctuations in blood glucose due to diabetes. Furthermore,GLP-1 can reduce appetite and reduce weight. Obesity is an inherentconsequence of poor control of glucose metabolism and this only servesto aggravate the diabetic condition.

Clinical application of natural GLP-1 is limited because it is rapidlydegraded in the circulation (half-life is several minutes). To maintaintherapeutic levels in the circulation requires constant administrationof high doses using pumps or patch devices which adds to the cost oftreatment. This is inconvenient for long term chronic use especially inconjunction with all the other medications for treating diabetes andmonitoring of glucose levels. The GLP-1 fusion proteins retain theactivity of GLP-1 but have the long half-life (14-17 days), solubility,and biodistribution properties of transferrin (mTf) and they can beadministered orally. These properties could provide for a low cost,small volume, monthly s.c. (subcutaneous) injection and this type ofproduct is absolutely needed for long term chronic use.

Insulin

Human insulin contains two peptide chains, known as the A and B chains,which are 21 and 30 amino acids in length, respectively, and which areconnected by two cystine disulphide bridges. This peptide has amolecular weight of approximately 6 kDa. The immediate precursor ofinsulin is proinsulin, a single chain peptide composed of the B and Achains linked to a connecting peptide of approximately 31 amino acids,known as the C-peptide, by adjacent pairs of basic residues. Thearrangement of these three peptides in the proinsulin molecule,beginning with the amino-terminal end, is as follows: Bchain-Arg-Arg-C-peptide-Lys-Arg-A chain. When translated into mRNA,however, preproinsulin is produced, which contains proinsulin joined atits amino-terminal end to a largely hydrophobic signal peptide 24 aminoacids in length.

Preproinsulin is synthesized in pancreatic beta cells located within theislets of Langerhans, which are dispersed throughout the pancreas.Removal of the signal peptide occurs in the rough endoplasmic reticulum,and the resulting proinsulin is then transported to the Golgi apparatusfor packaging into secretion granules. The folded proinsulin isstabilized by disulfide bonds. During processing of the secretiongranules, the folded proinsulin molecule is cleaved by specificproteases at the paired basic residues to liberate insulin and theC-peptide.

Diabetes mellitus is a disease that affects approximately 17 millionpeople in the United States, or 6.2% of the population. About onemillion people over the age of 20 are diagnosed with diabetes annually,and diabetes is the sixth leading cause of death in the United States(http://www.niddk.nih.gov/health/diabetes/pubs/dmstats/dmstats.htm#7).Approximately 90-95% of diabetes cases are Type 2, formerly calledadult-onset diabetes, which begins as insulin resistance (failure of thebody's cells to use insulin properly) as progresses to an inability ofthe pancreas to produce insulin. Type 1 diabetes, formerly calledjuvenile diabetes or insulin-dependent diabetes, accounts for theremaining 5-10% of diabetes cases. In type 1 diabetes, the body's immunesystem destroys the beta cells in the pancreas, which manufactureinsulin.

Because human insulin contains only 51 amino acid residues, it isreadily made by recombinant techniques, and a large number of insulinanalogues and variants have been prepared. Any of these analogues orvariants can be used to make mTf fusion proteins of the invention.

Diabetics typically require insulin replacement therapy, which involvesone or more doses of the drug per day by subcutaneous injection.Treatment by injection, however, is both psychologically and physicallypainful, as well as demanding of technical expertise, and many diabeticsrequire assistance in administering injections. Oral formulations ofinsulin have not been successful, however, because the peptide israpidly degraded in the acidic environment of the GI tract, particularlyin the stomach. Nevertheless, alternatives to injection, such as oral,nasal and topical formulations have been attempted. U.S. Pat. No.5,824,638, Burnside et al., describes oral emulsion preparations inwhich insulin is dissolved in a hydrophilic phase, such as water, salineor a water-miscible alcohol, and dispersed with a surfactant in ahydrophobic phase, such as a long chain fatty acid or fatty acid ester.Although an emulsion keeps insulin dispersed, it cannot protect thepeptide from the harsh conditions of the stomach. Nasal preparations,which deliver insulin in an aerosol to the lungs, are disclosed in U.S.Pat. No. 6,427,681, Gonda et al., while topical preparations aredisclosed in U.S. Pat. No. 6,399,566, Dardai et al.

Modified insulins for injection, containing amino acid substitutions orglycosylated residues, to enhance activity, inhibit degradation orinhibit peptide aggregation have also been developed (see U.S. Pat. No.4,478,746, Kim et al., glycosylated insulin derivatives; U.S. Pat. No.4,992,418, Katsoyanis et al., Asp₁₀-containing insulin (B chain) forincreased activity; U.S. Pat. No. 5,716,927, Balschmidt et al., Lys orArg at position 28 in the B chain, or A18, A21 or B3 modified from Asn,or other modifications at the C-terminal end of the B chain, to preventaggregation and reduced activity) Additional amino acid substitutionsthat confer a longer active phase, because they can be acylated, aredisclosed in U.S. Pat. No. 5,750,497, Havelund et al. A21 B3 and B30 canbe replaced by any amino acid except Lys, Arg or Cys. B1 may be deleted,and B30 may be replaced by a lipophilic chain of 10-24 carbon atoms.Fusion proteins for improved recombinant production of insulin (higheryields, soluble, allowing correct folding) are described in U.S. Pat.No. 6,534,288, Habermann et al. These peptides contain a fusion portionat the amino terminal end of the B chain, followed by amino acidsRDVP-Y_(n)-A chain, where Y is a peptide 2-50 amino acids in length,terminating with a basic amino acid.

The present invention includes fusion proteins comprising transferrinand an insulin protein or peptide. In one embodiment, the fusionproteins are formulated for oral delivery. The present invention,therefore, also includes methods of orally administering insulin fusionproteins of the invention to a patient in need thereof, in particular, adiabetic patient.

In one embodiment, the present invention includes transferrin fusionprotein comprising single chain insulin analog (Lee et al., 2000,Nature, 408: 483). In another embodiment, the insulin in the transferrinfusion protein may contain a protease cleavage site specific to thegastrointestinal (GI) tract, or a specific part of the gastrointestinaltract, such that the site would be recognized by one or more enzymes inthe GI tract. The proinsulin could be activated in this manner. Thecleavage site could reside in the peptide linking the A and B chain.

EPO Mimetic Peptide (EMP)

Erythropoietin (EPO) is a glycoprotein hormone that is synthesized inthe kidneys of mammals for stimulating mitotic cell division anddifferentiation of erythrocyte precursor cells. Accordingly, EPO acts tostimulate and regulate the production of erythrocytes. Because of itsrole in red blood cell formation, EPO is useful in both the diagnosisand the treatment of blood disorders characterized by low or defectivered blood cell production.

Studies have shown the efficacy of EPO therapy in a variety of diseasestates, disorders, and states of hematologic irregularity, for example,beta-thalassemia (Vedovato et al. (1984) Acta. Haematol. 71:211-213);cystic fibrosis (Vichinsky et al. (1984) J. Pediatric 105:15-21);pregnancy and menstrual disorders (Cotes et al. (1983) Brit. J. Ostet.Gyneacol. 90:304-311); early anemia of prematurity (Haga et al. (1983)Acta Pediatr. Scand. 72:827-831); spinal cord injury (Claus-Walker etal. (1984) Arch. Phys. Med. Rehabil. 65:370-374); space flight (Dunn etal. (1984) Eur. J. Appl. Physiol. 52:178-182); acute blood loss (Milleret al. (1982) Brit. J. Haematol. 52:545-590); aging (Udupa et al. (1984)J. Lab. Clin. Med. 103:574-588); various neoplastic disease statesaccompanied by abnormal erythropoiesis (Dainiak et al. (1983) Cancer5:1101-1106); and renal insufficiency (Eschbach et al. (1987) N. Eng. J.Med. 316:73-78). During the last fifteen years, EPO has been used forthe treatment of the anemia of renal failure, anemia of chronic diseaseassociated with rheumatoid arthritis, inflammatory bowel disease, AIDS,and cancer, as well as for the treatment of anemia in hematopoieticmalignancies, post-bone marrow transplantation, and autologous blooddonation.

The activity of EPO is mediated by its receptor. The EPO-receptor(EPO-R) belongs to the class of growth-factor-type receptors which areactivated by a ligand-induced protein dimerization. Other hormones andcytokines such as human growth hormone (hGH), granulocyte colonystimulating factor (G-CSF), epidermal growth factor (EGF) and insulincan cross-link two receptors resulting in juxtaposition of twocytoplasmic tails. Many of these dimerization-activated receptors haveprotein kinase domains within the cytoplasmic tails that phosphorylatethe neighboring tail upon dimerization. While some cytoplasmic tailslack intrinsic kinase activity, these function by association withprotein kinases. The EPO receptor is of the latter type. In each case,phosphorylation results in the activation of a signaling pathway.

There has been an increasing interest in molecular mimicry with EPOpotency. For example, dimerization of the erythropoietin receptor (EPOR)in the presence of either natural EPO or synthetic EPO mimetic peptides(EMPs) is the extracellular event that leads to activation of thereceptor and downstream signal transduction events. In general, there isan interest in obtaining mimetics with equivalent potency to EPO.

Wrighton et al (1996, Science, 273:458-463) employed phage display whererandom peptides are to be exposed on coat proteins of filamentous phage.A library of random peptide-phage was allowed to bind to andsubsequently eluted from the extracellular domain of EPO receptor in thescreening system. They used weak-binding system to first fish out EPOdomain-weak-binding (Kd 10 mM) CRIGPITWVC (SEQ ID NO: 10) as theconsensus sequence. Consequently, a 20-amino acid peptide, EMP1,(GGTYSCHFGPLTWVCKPQGG, SEQ ID NO: 11) with an affinity (Kd) of 200 nM,compared to 200 pM for EPO was isolated, the sequence of which does notactually exist in the native EPO. The crystal structure at 2.8 Åresolution of a complex of this mimetic agonist peptide with theextracellular domain of EPO receptor revealed that a peptide dimerinduces an almost perfect twofold dimerization of the receptor (Livnahet al., 1996 Science, 273 (274): 464-471). This 20-amino acid peptidehas a b-sheet structure and is stabilized by the C—C disulfide bond.

The biological activity of EMP1 indicates that EMP1 can act as an EPOmimetic. For example, EMP1 competes with EPO in receptor binding assaysto cause cellular proliferation of cell lines engineered to beresponsive to EPO (Wrighton et al., 1996, Science, 273:458-463). BothEPO and EMP1 induce a similar cascade of phosphorylation events and cellcycle progression in EPO responsive cells (Wrighton et al., 1996,Science, 273:458-463). Further, EMP1 demonstrates significanterythropoietic effects in mice as monitored by two different in vivoassays of nascent red blood cell production (Wrighton et al., 1996,Science, 273:458-463).

Johnson et al. (1998, Biochemistry, 37:3699-3710) identified the minimalpeptide that retained activity in the assays for EPO mimetic action.Using N- and C-terminal deletions, they found that the minimal activepeptide is EMP20 having the sequence, YSCHFGPLTWVCK (SEQ ID NO: 12),namely amino acids 4 through 16 of EMP1. They also found Tyr4 and Trp 13of EMP1 are critical for mimetic action.

The present invention provides EMP1/transferrin fusion proteins withincreased half-life and pharmaceutical compositions comprising suchfusion proteins. Any EMP sequence may be used to make EMP1/transferrinfusion proteins, including EMP1 sequences wherein one or more C residuesis deleted or replaced. These sequences can then be inserted into a mTfloop to provide three dimensional structure to the EMP1 region of thefusion protein. The present invention contemplates the use of the fusionprotein to treat various diseases and conditions associated with EPOsuch as but not limited to those described above.

In one embodiment of the present invention, the pharmaceuticalcompositions comprising the EMP1/transferrin fusion protein and may beformulated by any of the established methods of formulatingpharmaceutical compositions, e.g. as described in Remington'sPharmaceutical Sciences, 1985. The composition may be in a form suitedfor systemic injection or infusion and may, as such, be formulated witha suitable liquid vehicle such as sterile water or an isotonic saline orglucose solution. These pharmaceutical compositions may contain buffers,salts and other excipients to stabilize the composition or assist in thedelivery of the transferrin fusion proteins.

In a preferred embodiment, the present invention provides a method fortreating disorders associated with EPO. The method is accomplished byadministering a EMP1/transferrin fusion protein provided herein for atime and under conditions sufficient to alleviate the symptoms of thedisorder, i.e. sufficient to effect dimerization or biologicalactivation of EPO receptors. In the case of EPO such methodology isuseful in the treatment of end-stage renal failure/dialysis; anemia,especially associated with AIDS or chronic inflammatory diseases such asrheumatoid arthritis and chronic bowel inflammation; auto-immunedisease; and for boosting the red blood cell count of patient whennecessary, e.g. prior to surgery or as pretreatment to transfusion. TheEMP1/transferrin fusion protein of the present invention which behave asEPO agonists can be used to activate megakaryocytes.

Since EPO has been shown to have a mitogenic and chemotactic effect onvascular endothelial cells as well as an effect on central cholinergicneurons (Amagnostou et al. (1990) Proc. Natl. Acad. Sci. USA87:597805982; Konishi et al. (1993) Brain Res. 609:29-35), the compoundsof this invention can also be used to treat a variety of vasculardisorders, such as promoting wound healing, growth of collateralcoronary blood vessels (such as those that may occur after myocardialinfarction), trauma, and post vascular graft treatment, and a variety ofneurological disorders, generally characterized by low absolute levelsof acetyl choline or low relative levels of acetyl choline as comparedto other neuroactive substances e.g., neurotransmitters.

Accordingly, the present invention includes pharmaceutical compositionscomprising, as an active ingredient, the EMP1/transferrin fusion proteinof the present invention in association with a pharmaceutical carrier ordiluent. The EMP1/transferrin fusion protein of this invention can beadministered by oral, parenteral (intramuscular, intraperitoneal,intravenous (IV) or subcutaneous injection), transdermal (eitherpassively or using iontophoresis or electroporation) or transmucosal(nasal, vaginal, rectal, or sublingual) routes of administration indosage forms appropriate for each route of administration.

Solid dosage forms for oral administration include capsules, tablets,pill, powders, and granules. In such solid dosage forms, the activecompound is admixed with at least one inert pharmaceutically acceptablecarrier such as sucrose, lactose, or starch. Such dosage forms can alsocomprise, as it normal practice, additional substances other than inertdiluents, e.g., lubricating, agents such as magnesium stearate. In thecase of capsules, tablets and pills, the dosage forms may also comprisebuffering, agents. Tablets and pills can additionally be prepared withenteric coatings.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, with the elixirscontaining inert diluents commonly used in the art, such as water.Besides such inert diluents, compositions can also include adjuvants,such as wetting agents, emulsifying and suspending agents, andsweetening, flavoring and perfuming agents.

Preparations according to this invention for parenteral administrationinclude sterile aqueous or non-aqueous solutions, suspensions, oremulsions. Examples of non-aqueous solvents or vehicles are propyleneglycol, polyethylene glycol, vegetable oils, such as olive oil and cornoil, gelatin, and injectable organic esters such as ethyl oleate. Suchdosage forms may also contain adjuvants such as preserving, wetting,emulsifying, and dispersing agents. They may be sterilized by, forexample, filtration through a bacteria retaining filter, byincorporating sterilizing agents into the compositions, by irradiatingthe compositions, or by heating the compositions. They can also bemanufactured using sterile water, or some other sterile injectablemedium, immediately before use.

Compositions for rectal or vaginal administration are preferablysuppositories which may contain, in addition to the active substance,excipients such as cocoa butter or a suppository wax. Compositions fornasal or sublingual administration are also prepared with standardexcipients well known in the art.

The dosage of active ingredient in the compositions of this inventionmay be varied; however, it is necessary that the amount of the activeingredient shall be such that a suitable dosage form is obtained. Theselected dosage depends upon the desired therapeutic effect, on theroute of administration, and on the duration of the treatment desired.Generally dosage levels of between 0.001 to 10 mg/kg of body weightdaily are administered to mammals.

Moreover, the present invention also contemplates the use of thetransferrin fusion protein comprising EMP1 or analogs thereof for themanufacture of a medicinal product which can be used in the treatment ofdiseases associated with low or defective red blood cell production.Examples of such diseases are not limited to those described above.

T-20 and T-1249

HIV infection is pandemic and HIV associated diseases represent a majorworld health problem. Although considerable effort is being put into thesuccessful design of effective therapeutics, currently no curativeanti-retroviral drugs against AIDS exist. In attempts to develop suchdrugs, several stages of the HIV life cycle have been considered astargets for therapeutic intervention (Mitsuya, H. et al., 1991, FASEB J.5:2369-2381). For example, virally encoded reverse transcriptase hasbeen one focus of drug development. A number ofreverse-transcriptase-targeted drugs, including 2′,3′-dideoxynucleosideanalogs such as AZT, ddI, ddc, and d4T have been developed which havebeen shown to been active against HIV (Mitsuya, H. et al., 1991, Science249:1533-1544). While beneficial, these nucleoside analogs are notcurative, probably due to the rapid appearance of drug resistant HIVmutants (Lander, B. et al., 1989, Science 243:1731-1734). In addition,the drugs often exhibit toxic side effects, such as bone marrowsuppression, vomiting, and liver function abnormalities.

Entry inhibitors are distinct from the existing classes of drugs thatfight HIV. Other drugs work inside the infected cell. Nucleoside reversetranscriptase inhibitors such as AZT and abacavir and non-nucleosidereverse transcriptase inhibitors like nevirapine and efavirenz all actby shutting down the reverse transcriptase enzyme that HIV uses toreplicate itself once it is inside the cell. Protease inhibitors shutdown the viral protease enzyme HIV uses to package itself up for export.By contrast, entry inhibitors are drugs that interfere with theprocesses involved in the virus' initial assault on the cell's outermembrane.

T-20 is the most studied of all the entry inhibitors and is the firstmember of the fusion inhibitor class. Unlike existing AIDS drugs thatwork inside the cell and target viral enzymes involved in thereplication of the virus, T-20 inhibits fusion of HIV with host cellsbefore the virus enters the cell and begins its replication process.T-20 binds to one of the two helical domains of gp41. Gp41 is aspring-loaded HIV-1 protein that is activated when CD4 binds to HIVgp-120. The fusion action of gp41 is inhibited if its two helicaldomains cannot fold together. T-20 binds to gp41, effectively keepingthe protein from functioning. It has been shown in early, single-armclinical studies to be about as potent as a protease inhibitor byitself-giving greater than 10 fold reductions in viral load and to besafe in combination with other antiretrovirals.

U.S. Pat. No. 5,464,933 discloses T-20 (pentafuside, DP-178) as a 36amino acid synthetic peptide. Since this drug is a peptide, it cannot begiven orally because it is readily broken down by the digestive system.When administered by subcutaneous injection, T-20 achieves sufficientlevels in the blood to have anti-HIV activity. It is administered bysubcutaneous injection twice daily. However, patients develop skinreactions at the injection site. The most frequently reported treatmentrelated adverse events were mild to moderate local injection sitereactions. These consist of mild pain, temporary swelling and redness atthe site of injection.

U.S. Pat. No. 6,479,055 discloses peptide analogs of the DP-178(peptides corresponding to amino acid residues 638 to 673 oftransmembrane protein gp41 of HIV-1_(LAI), which exhibit anti-membranefusion capability, antiviral activity, such as the ability to inhibitHIV transmission to uninfected CD-4⁺ cells, or an ability to modulateintracellular processes involving coiled-coil peptide structures.Further, the patent relates to the use of DP-178 and DP-178 portionsand/or analogs as antifusogenic or antiviral compounds or as inhibitorsof intracellular events involving coiled-coil peptide structures.Further, the patent teaches the use of the peptides as diagnosticagents. For example, a DP178 peptide may be used as an HIVsubtype-specific diagnostic.

T-1249 is a sister compound of T-20. Like T-20, T-1249 targets the HIVglycoprotein known as gp41 which HIV uses to bind onto CD4 cells. T-1249has shown potent anti-HIV effects in animal and laboratory studies.Preliminary safety, dosing and efficacy studies in humans have providedsupport for ongoing research.

T-1249 is currently administered by subcutaneous (under the skin)injection once or twice daily. The first safety study of T-1249conducted in humans found two serious adverse events: hypersensitivityreaction (oral ulcers, maculopapular rash, fever) and severeneutropenia. Forty percent of recipients developed injection sitereactions but these were deemed to be mild. Dizziness, diarrhea,headache and fever have also been reported by recipients. Nodose-limiting toxicity was identified and experiments with higher dosesare likely.

T-1249 has completed phase I/II safety and dosing studies. Initialresults indicated that higher doses produced an average viral load dropof 1.3 log.

Dose-dependent decreases in HIV RNA have been reported. In the study ofT-1249, the average reduction from baseline ranged from 0.29 to 1.96 logcopies/ml (Gulick 2002).

The present invention provides transferrin fusion proteins comprisingT-20, T-1249, or analogs thereof with increased half-life andpharmaceutical compositions comprising such fusion proteins. The presentinvention also provides pharmaceutical compositions comprising thesetransferrin fusion proteins for therapeutic purposes. The presentinvention contemplates the use of such fusion proteins as inhibitors ofhuman and non-human retroviral, especially HIV, transmission touninfected cells. The human retroviruses whose transmission may beinhibited by the peptides of the invention include, but are not limitedto all strains of HIV-1 and HIV-2 and the human T-lymphocyte viruses(HTLV-I, II, III). The non-human retroviruses whose transmission may beinhibited by the peptides of the invention include, but are not limitedto bovine leukosis virus, feline sarcoma and leukemia viruses, simiansarcoma and leukemia viruses, and sheep progress pneumonia viruses.

With respect to HIV, the transferrin fusion protein of the presentinvention comprising T-20, T-1249 or analogs thereof may be used as atherapeutic in the treatment of AIDS. These transferrin fusion proteinsmay be administered using techniques well known to those in the art.Preferably, the pharmaceutical compositions comprising these transferrinfusion proteins are formulated and administered systemically. Techniquesfor formulation and administration may be found in “Remington'sPharmaceutical Sciences” 18th ed., 1990 Mack Publishing Co., Easton, Pa.Suitable routes may include oral, rectal, transmucosal, or intestinaladministration; parenteral delivery, including intramuscular,subcutaneous, intramedullary injections, as well as intrathecal, directintraventricular, intravenous, intraperitoneal, intranasal, orintraocular injections, just to name a few. Most preferably,administration is intravenous. For injection, the transferrin fusionproteins comprising T-20, T1249, or analogs thereof may be formulated inaqueous solutions, preferably in physiologically compatible buffers suchas Hanks' solution, Ringer's solution, or physiological saline buffer.For such transmucosal administration, penetrants appropriate to thebarrier to be permeated are used in the formulation. Such penetrants aregenerally known in the art.

In addition, the transferrin fusion protein comprising T-20, T1249, oranalogs thereof may be used as a prophylactic measure in previouslyuninfected individuals after acute exposure to an HIV virus. Examples ofsuch prophylactic use of the peptides may include, but are not limitedto, prevention of virus transmission from mother to infant and othersettings where the likelihood of HIV transmission exists, such as, forexample, accidents in health care settings wherein workers are exposedto HIV-containing blood products. The transferrin fusion proteins of thepresent invention comprising T-20, T-1249, or analogs thereof in suchcases may serve the role of a prophylactic vaccine, wherein the hostraises antibodies against the fusion proteins of the invention, whichthen serve to neutralize HIV viruses by, for example, inhibiting furtherHIV infection. Administration of the transferrin fusion proteins of theinvention as a prophylactic vaccine, therefore, would compriseadministering to a host a concentration of transferrin fusion proteineffective in raising an immune response which is sufficient toneutralize HIV, by, for example, inhibiting HIV ability to infect cells.The exact concentration will depend upon the specific peptide in thetransferrin fusion protein to be administered, but may be determined byusing standard techniques for assaying the development of an immuneresponse which are well known to those of ordinary skill in the art. Thetransferrin fusion proteins to be used as vaccines are usuallyadministered intramuscularly.

Effective dosages of the transferrin fusion proteins comprising T-20,T-1249, or analogs thereof to be administered may be determined throughprocedures well known to those in the art which address such parametersas biological half-life, bioavailability, and toxicity. Given the datapresented below in Section 6, DP-178, for example, may prove efficaciousin vivo at doses required achieve circulating levels of 10 ng per ml ofpeptide.

Furthermore, the present invention contemplates the use of thetransferrin fusion proteins comprising T-20, T-1249, or analogs thereoffor the manufacture of a medicinal product for the treatment of diseasesassociated with the transmission of a virus.

Delivery of a Drug or Therapeutic Protein to the Inside of a Cell and/orAcross the Blood Brain Barrier (BBB)

Within the scope of the invention, the modified transferrin fusionproteins may be used as a carrier to deliver a molecule or smallmolecule therapeutic complexed to the ferric ion of transferrin to theinside of a cell or across the blood brain barrier or other barriersincluding across the cell membrane of any cell type that naturally orengineered to express a Tf receptor. In these embodiments, the Tf fusionprotein will typically be engineered or modified to inhibit, prevent orremove glycosylation to extend the serum half-life of the fusion proteinand/or therapeutic protein portion. The addition of a targeting peptideis specifically contemplated to further target the Tf fusion protein toa particular cell type, e.g., a cancer cell.

In one embodiment, the iron-containing, anti-anemic drug,ferric-sorbitol-citrate complex is loaded onto a modified Tf fusionprotein of the invention. Ferric-sorbitol-citrate (FSC) has been shownto inhibit proliferation of various murine cancer cells in vitro andcause tumor regression in vivo, while not having any effect onproliferation of non-malignant cells (Poljak-Blazi et al. (June 2000)Cancer Biotherapy and Radiopharmaceuticals (United States),15/3:285-293).

In another embodiment, the antineoplastic drug Adriamycin® (doxorubicin)and/or the chemotherapeutic drug bleomycin, both of which are known toform complexes with ferric ion, is loaded onto a Tf fusion protein ofthe invention. In other embodiments, a salt of a drug, for instance, acitrate or carbonate salt, may be prepared and complexed with the ferriciron that is then bound to Tf. As tumor cells often display a higherturnover rate for iron; transferrin modified to carry at least oneanti-tumor agent, may provide a means of increasing gent exposure orload to the tumor cells. (Demant, E. J., (1983) Eur. J. Biochem.137/(1-2):113-118; Padbury et al. (1985) J. Biol. Chem.260/13:7820-7823).

Pharmaceutical Formulations and Treatment Methods

The modified fusion proteins of the invention may be administered to apatient in need thereof using standard administration protocols. Forinstance, the modified Tf fusion proteins of the present invention canbe provided alone, or in combination, or in sequential combination withother agents that modulate a particular pathological process. As usedherein, two agents are said to be administered in combination when thetwo agents are administered simultaneously or are administeredindependently in a fashion such that the agents will act at the same ornear the same time.

The agents of the present invention can be administered via parenteral,subcutaneous, intravenous, intramuscular, intraperitoneal, transdermaland buccal routes. For example, an agent may be administered locally toa site of injury via microinfusion. Alternatively, or concurrently,administration may be noninvasive by either the oral, inhalation, nasal,or pulmonary route. The dosage administered will be dependent upon theage, health, and weight of the recipient, kind of concurrent treatment,if any, frequency of treatment, and the nature of the effect desired.

While any method of administration may be used to deliver the mTF fusionproteins of the invention, administration or delivery orally may be apreferred embodiment for certain classes of fusion proteins or to treatcertain conditions.

The present invention further provides compositions containing one ormore fusion proteins of the invention. While individual needs vary,determination of optimal ranges of effective amounts of each componentis within the skill of the art. Typical dosages comprise about 1 pg/kgto about 100 mg/kg body weight. The preferred dosages for systemicadministration comprise about 100 ng/kg to about 100 mg/kg body weight.The preferred dosages for direct administration to a site viamicroinfusion comprise about 1 ng/kg to about 1 mg/kg body weight. Whenadministered via direct injection or microinfusion, modified fusionproteins of the invention may be engineered to exhibit reduced or nobinding of iron to prevent, in part, localized iron toxicity.

In addition to the pharmacologically active fusion protein, thecompositions of the present invention may contain suitablepharmaceutically acceptable carriers comprising excipients andauxiliaries that facilitate processing of the active compounds intopreparations which can be used pharmaceutically for delivery to the siteof action. Suitable formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form, forexample, water-soluble salts. In addition, suspensions of the activecompounds as appropriate oily injection suspensions may be administered.Suitable lipophilic solvents or vehicles include fatty oils, forexample, sesame oil, or synthetic fatty acid esters, for example, ethyloleate or triglycerides. Aqueous injection suspensions may containsubstances which increase the viscosity of the suspension including, forexample, sodium carboxymethyl cellulose, sorbitol and dextran.Optionally, the suspension may also contain stabilizers. Liposomes canalso be used to encapsulate the agent for delivery into the cell.

The pharmaceutical formulation for systemic administration according tothe invention may be formulated for enteral, parenteral or topicaladministration. Indeed, all three types of formulations may be usedsimultaneously to achieve systemic administration of the activeingredient. Suitable formulations for oral administration include hardor soft gelatin capsules, pills, tablets, including coated tablets,elixirs, suspensions, syrups or inhalations and controlled release formsthereof.

In practicing the methods of this invention, the agents of thisinvention may be used alone or in combination, or in combination withother therapeutic or diagnostic agents. In certain preferredembodiments, the compounds of this invention may be co-administeredalong with other compounds typically prescribed for these conditionsaccording to generally accepted medical practice. The compounds of thisinvention can be utilized in vivo, ordinarily in mammals, such ashumans, sheep, horses, cattle, pigs, dogs, cats, rats and mice, ex vivoor in vitro.

Modified fusion proteins of the present invention may be used in thediagnosis, prognosis, prevention and/or treatment of diseases and/ordisorders relating to diseases and disorders of the endocrine system,the nervous system, the immune system, respiratory system,cardiovascular system, reproductive system, digestive system, diseasesand/or disorders relating to cell proliferation, and/or diseases ordisorders relating to the blood.

In yet other embodiments of the invention, modified Tf fusion proteinsmay be used in the diagnosis, prognosis, prevention and/or treatment ofdiseases and/or disorders relating to diseases and disorders known to beassociated with or treatable by therapeutic protein moieties as known inthe art and exemplified by PCT Patent Publication Nos. WO 01/79258, WO01/77137, WO 01/79442, WO 01/79443, WO 01/79444 and WO 01/79480, all ofwhich are herein incorporated by reference in their entirety.Accordingly, the present invention encompasses a method of treating adisease or disorder listed in the “Preferred Indication Y” column ofTable 1 comprising administering to a patient in which such treatment,prevention or amelioration is desired a modified transferrin fusionprotein of the invention that comprises a therapeutic protein portioncorresponding to a therapeutic protein disclosed in the “TherapeuticProtein X” column of Table 1 in an amount effective to treat, prevent orameliorate the disease or disorder.

In certain embodiments, a transferrin fusion protein of the presentinvention may be used to diagnose and/or prognose diseases and/ordisorders.

Modified transferrin fusion proteins of the invention andpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in treating, preventing, diagnosing and/or prognosingdiseases, disorders, and/or conditions of the immune system. Moreover,fusion proteins of the invention and/or polynucleotides encodingtransferrin fusion proteins of the invention can be used as a marker ordetector of a particular immune system disease or disorder.

In a preferred embodiment fusion proteins of the invention and/orpolynucleotides encoding modified transferrin fusion proteins of theinvention could be used as an agent to boost immunoresponsiveness amongimmunodeficient individuals. In specific embodiments, fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention could be used as an agent to boostimmunoresponsiveness among B cell and/or T cell immunodeficientindividuals.

The modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in treating, preventing, diagnosing, and/or prognosingautoimmune disorders. Many autoimmune disorders result frominappropriate recognition of self as foreign material by immune cells.This inappropriate recognition results in an immune response leading tothe destruction of the host tissue. Therefore, the administration offusion proteins of the invention and/or polynucleotides encodingtransferrin fusion proteins of the invention, that can inhibit an immuneresponse, particularly the proliferation, differentiation, or chemotaxisof T-cells, may be an effective therapy in preventing autoimmunedisorders.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in treating, preventing, prognosing, and/or diagnosingdiseases, disorders, and/or conditions of hematopoietic cells.Transferrin fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention could be used toincrease differentiation and proliferation of hematopoietic cells,including the pluripotent stem cells, in an effort to treat or preventthose diseases, disorders, and/or conditions associated with a decreasein certain (or many) types hematopoietic cells, including but notlimited to, leukopenia, neutropenia, anemia, and thrombocytopenia.

Alternatively, modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventioncould be used to increase differentiation and proliferation ofhematopoietic cells, including the pluripotent stem cells, in an effortto treat or prevent those diseases, disorders, and/or conditionsassociated with an increase in certain (or many) types of hematopoieticcells, including but not limited to, histiocytosis.

Allergic reactions and conditions, such as asthma (particularly allergicasthma) or other respiratory problems, may also be treated, prevented,diagnosed and/or prognosing and using modified fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention. Moreover, these molecules can be used to treat, prevent,prognose, and/or diagnose anaphylaxis, hypersensitivity to an antigenicmolecule, or blood group incompatibility.

Additionally, modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the invention,may be used to treat, prevent, diagnose and/or prognose IgE-mediatedallergic reactions. Such allergic reactions include, but are not limitedto, asthma, rhinitis, and eczema. In specific embodiments, fusionproteins of the invention and/or polynucleotides encoding transferrinfusion proteins of the invention may be used to modulate IgEconcentrations in vitro or in vivo.

Moreover, modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionhave uses in the diagnosis, prognosis, prevention, and/or treatment ofinflammatory conditions. For example, since fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may inhibit the activation, proliferation, and/ordifferentiation of cells involved in an inflammatory response, thesemolecules can be used to prevent and/or treat chronic and acuteinflammatory conditions. Such inflammatory conditions include, but arenot limited to, for example, inflammation associated with infection(e.g., septic shock, sepsis, or systemic inflammatory responsesyndrome), ischemia-reperfusion injury, endotoxin lethality,complement-mediated hyperacute rejection, nephritis, cytokine orchemokine induced lung injury, inflammatory bowel disease, Crohn'sdisease, over production of cytokines (e.g., TNF or IL-1.), respiratorydisorders (e.g., asthma and allergy); gastrointestinal disorders (e.g.,inflammatory bowel disease); cancers (e.g., gastric, ovarian, lung,bladder, liver, and breast); CNS disorders (e.g., multiple sclerosis;ischemic brain injury and/or stroke, traumatic brain injury),neurodegenerative disorders (e.g., Parkinson's disease and Alzheizmer'sdisease); AIDS-related dementia; and prion disease); cardiovasculardisorders (e.g., atherosclerosis, myocarditis, cardiovascular disease,and cardiopulmonary bypass complications); as well as many additionaldiseases, conditions, and disorders that are characterized byinflammation (e.g., hepatitis, rheumatoid arthritis, gout, trauma,pancreatitis, sarcoidosis, dermatitis, renal ischemia-reperfusioninjury, Grave's disease, systemic lupus erythematosus, diabetesmellitus, and allogenic transplant rejection).

Because inflammation is a fundamental defense mechanism, inflammatorydisorders can affect virtually any tissue of the body. Accordingly,modified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention, have uses in thetreatment of tissue-specific inflammatory disorders, including, but notlimited to, adrenalitis, alveolitis, angiocholecystitis, appendicitis,balanitis, blepharitis, bronchitis, bursitis, carditis, cellulitis,cervicitis, cholecystitis, chorditis, cochiftis, colitis,conjunctivitis, cystitis, dermatitis, diverticulitis, encephalitis,endocarditis, esophagitis, eustachitis, fibrositis, folliculitis,gastritis, gastroenteritis, gingivitis, glossitis, hepatosplenitis,keratitis, labyrinthitis, laryngitis, lymphangitis, mastitis, mediaotitis, meningitis, metritis, mucitis, myocarditis, myosititis,myringitis, nephritis, neuritis, orchitis, osteochondritis, otitis,pericarditis, peritendonitis, peritonitis, pharyngitis, phlebitis,poliomyelitis, prostatititis, Pulpitis, retinitis, rhinitis,salpingitis, scleritis, sclerochoroiditis, scrotitis, sinusitis,spondylitis, steatitis, stomatitis, synovitis, syringitis, tendonitis,tonsillitis, urethritis, and vaginitis.

In specific embodiments, modified fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention, are useful to diagnose, prognose, prevent, and/or treat organtransplant rejections and graft-versus-host disease (GVHD). Organrejection occurs by host immune cell destruction of the transplantedtissue through an immune response. Similarly, an immune response is alsoinvolved in GVHD, but, in this case, the foreign transplanted immunecells destroy the host tissues. Polypeptides, antibodies, orpolynucleotides of the invention, and/or agonists or antagoniststhereof, that inhibit an immune response, particularly the activation,proliferation, differentiation, or chemotaxis of T-cells, may be aneffective therapy in preventing organ rejection or GVHD.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used as an adjuvant to enhance anti-viralimmune responses. Anti-viral immune responses that may be enhanced usingthe compositions of the invention as an adjuvant include virus and virusassociated diseases or symptoms described herein or otherwise known inthe art. In specific embodiments, the compositions of the invention areused as an adjuvant to enhance an immune response to a virus, disease,or symptom selected from the group consisting of AIDS, meningitis,Dengue, EBV, and hepatitis (e.g., hepatitis B). In another specificembodiment, the compositions of the invention are used as an adjuvant toenhance an immune response to a virus, disease, or symptom selected fromthe group consisting of: HIV/AIDS, respiratory syncytial virus, Dengue,rotavirus, Japanese B encephalitis, influenza A and B, parainfluenza,measles, cytomegalovirus, rabies, Junin, Chikungunya, Rift Valley Fever,herpes simplex, and yellow fever.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used as an adjuvant to enhanceanti-bacterial or anti-fungal immune responses. Anti-bacterial oranti-fungal immune responses that may be enhanced using the compositionsof the invention as an adjuvant include bacteria or fungus and bacteriaor fungus associated diseases or symptoms described herein or otherwiseknown in the art. In specific embodiments, the compositions of theinvention are used as an adjuvant to enhance an immune response to abacterium or fungus disease, or symptom selected from the groupconsisting of tetanus, Diphtheria, botulism, meningitis type B, andcandidiasis.

In another specific embodiment, the compositions of the invention areused as an adjuvant to enhance an immune response to a bacterium orfungus, disease, or symptom selected from the group consisting of Vibriocholerae, Mycobacterium leprae, Salmonella typhi, Salmonella paratyphi,Neisseria meningitidis, Streptococcus pneumoniae, Group B streptococcus,Shigella spp., Enterotoxigenic Escherichia coli, Enterohemorrhagic E.coli, Borrelia burgdorferi, and Candida.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used as an adjuvant to enhanceanti-parasitic immune responses. Anti-parasitic immune responses thatmay be enhanced using the compositions of the invention as an adjuvantinclude parasite and parasite associated diseases or symptoms describedherein or otherwise known in the art. In specific embodiments, thecompositions of the invention are used as an adjuvant to enhance animmune response to a parasite. In another specific embodiment, thecompositions of the invention are used as an, adjuvant to enhance animmune response to Plasmodium (malaria) or Leishmania.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention may also be employed to treat infectiousdiseases including silicosis, sarcoidosis, and idiopathic pulmonaryfibrosis; for example, by preventing the recruitment and activation ofmononuclear phagocytes.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used as an antigen for the generation ofantibodies to inhibit or enhance immune mediated responses againstpolypeptides of the invention.

In one embodiment, modified transferrin fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention are administered to an animal (e.g., mouse, rat, rabbit,hamster, guinea pig, pigs, micro-pig, chicken, camel, goat, horse, cow,sheep, dog, cat non-human primate, and human, most preferably human) toboost the immune system to produce increased quantities, of one or moreantibodies (e.g., IgG, IgA, IgM, and IgE), to induce higher affinityantibody production and immunoglobulin class switching (e.g., IgG, IgA,IgM, and IgE), and/or to increase an immune response.

In another embodiment, modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention are used in one or more of the applications describedherein, as they may apply to veterinary medicine.

In another specific embodiment, modified transferrin fusion proteins ofthe invention, and/or polynucleotides encoding transferrin fusionproteins of the invention are used as a means of blocking variousaspects of immune responses to foreign agents or self. Examples ofdiseases or conditions in which blocking of certain aspects of immuneresponses may be desired include autoimmune disorders such as lupus, andarthritis, as well as immunoresponsiveness to skin allergies,inflammation-, bowel disease, injury, and diseases/disorders associatedwith pathogens.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used as a therapy for preventing the Bcell proliferation and Ig secretion associated with autoimmune diseasessuch as idiopathic thrombocytopenic purpura, systemic lupuserythematosus and multiple sclerosis.

In another specific embodiment, modified transferrin fusion proteins orpolynucleotides encoding transferrin fusion proteins of the inventionare used as an inhibitor of B and/or T cell migration in endothelialcells. This activity disrupts tissue architecture or cognate responsesand is useful, for example in disrupting immune responses, and blockingsepsis.

In another specific embodiment, modified transferrin fusion proteins ofthe invention, and/or polynucleotides encoding transferrin fusionproteins of the invention are used as a therapy for chronichypergammaglobulinen evident in such diseases as monoclonal gammopathyof undetermined significance (MGUS), Waldenstrom's disease, relatedidiopathic monocional gammopathies, and plasmacytomas.

Another specific embodiment, modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be employed for instance to inhibit polypeptidechemotaxis and activation of macrophages and their precursors, and ofneutrophils, basophils, B lymphocytes and some T-cell subsets, e.g.,activated and CD8 cytotoxic T cells and natural killer cells, in certainautoimmune and chronic inflammatory and infective diseases. Examples ofautoimmune diseases are described herein and include multiple sclerosis,and insulin-dependent diabetes.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusion proteinof the invention may also be employed for treating atherosclerosis, forexample, by preventing monocyte infiltration in the artery wall.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or -polynucleotides encoding transferrin fusionproteins of the invention may be employed to treat adult respiratorydistress syndrome (ARDS).

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention may be useful for stimulating wound and tissuerepair, stimulating angiogenesis, and/or stimulating the repair ofvascular or lymphatic diseases or disorders. Additionally, fusionproteins of the invention and/or polynucleotides encoding transferrinfusion proteins of the invention may be used to stimulate theregeneration of mucosal surfaces.

In a specific embodiment, modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention are used to diagnose, prognose, treat, and/or prevent adisorder characterized by primary or acquired immunodeficiency,deficient serum immunoglobulin production, recurrent infections, and/orimmune system dysfunction. Moreover, modified fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be used to treat or prevent infections of the joints,bones, skin, and/or parotid glands, blood-borne infections (e.g.,sepsis, meningitis, septic arthritis, and/or osteomyelitis), autoimmunediseases (e.g., those disclosed herein), inflammatory disorders, andmalignancies, and/or any disease or disorder or condition associatedwith these infections, diseases, disorders and/or malignancies)including, but not limited to, Common Variable Immunodeficiency (CVID),other primary immune deficiencies, HIV disease, Chronic LymphocyticLeukemia (CLL), recurrent bronchitis, sinusitis, otitis media,conjunctivitis, pneumonia, hepatitis, meningitis, herpes zoster (e.g.,severe herpes zoster), and/or pneumocystis carnii. Other diseases anddisorders that may be prevented, diagnosed, prognosed, and/or treatedwith fusion proteins of the invention and/or polynucleotides encodingtransferrin fusion proteins of the invention include, but are notlimited to, HIV infection, HTLV-BLV infection, lymphopenia, phagocytebactericidal dysfunction anemia, thrombocytopenia, and hemoglobinuria.

In a specific embodiment, modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be used to diagnose, prognose, prevent, and/or treatcancers or neoplasms including immune cell or immune tissue-relatedcancers or neoplasms. Examples of cancers or neoplasms that may beprevented, diagnosed, or treated by fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention include, but are not limited to, acute myelogenous leukemia,chronic myelogenous leukemia, Hodgkin's disease, non-Hodgkin's lymphoma,acute lymphocytic leukemia (ALL), chronic lymphocyte leukemia,plasmacytomas, multiple myeloma, Burkitt's lymphoma, EBV transformeddiseases, and/or diseases and disorders described in the sectionentitled “Hyperproliferative Disorders” elsewhere herein.

In another specific embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used as a therapy for decreasing cellularproliferation of Large B-cell Lymphomas.

In specific embodiments, the compositions of the invention are used asan agent to boost immunoresponsiveness among B cell immunodeficientindividuals, such as, for example, an individual who has undergone apartial or complete splenectomy.

The modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be used to modulate hemostatic (the stopping of bleeding) orthrombolytic (clot dissolving) activity. For example, by increasinghemostatic or thrombolytic activity, fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention could be used to treat or prevent blood coagulation diseases,disorders, and/or conditions (e.g., afibrinogenemia, factordeficiencies, hemophilia), blood platelet diseases, disorders, and/orconditions (e.g. thrombocytopenia), or wounds resulting from trauma,surgery, or other causes. Alternatively, fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention that can decrease hemostatic or thrombolytic activitycould be used to inhibit or dissolve clotting. These molecules could beimportant in the treatment or prevention of heart attacks (infarction),strokes, or scarring.

In specific embodiments, the modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be used to prevent diagnose, prognose, and/or treatthrombosis, arterial thrombosis, venous thrombosis, thromboembolism,pulmonary embolism, atherosclerosis, myocardial infarction, transientischemic attack, unstable angina. In specific embodiments, thetransferrin fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention maybe used for theprevention of occulsion of saphenous grafts, for reducing the risk ofperiprocedural thrombosis as might accompany angioplasty procedures, forreducing the risk of stroke in patients with atrial fibrillationincluding nonrheumatic atria fibrillation, for reducing the risk ofembolism associated with mechanical heart valves and/or mitral valvesdisease. Other uses for the modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention, include, but are not limited to, the prevention ofocclusions in extracorporeal devices (e.g., intravascular canals,vascular access shunts in hemodialysis patients, hemodialysis machines,and cardiopulmonary bypass machines).

In another embodiment, modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention, may be used to prevent, diagnose, prognose, and/or treatdiseases and disorders of the blood and/or blood forming organsassociated with the tissue(s) in which the polypeptide of the inventionis expressed.

The modified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention may be used tomodulate hematopoietic activity (the formation of blood cells). Forexample, the transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be used to increase the quantity of all or subsets of blood cells,such as, for example, erythrocytes, lymphocytes (B or T cells), myeloidcells (e.g., basophils, eosinophils, neutrophils, mast cells,macrophages) and platelets. The ability to decrease the quantity ofblood cells or subsets of blood cells may be useful in the prevention,detection, diagnosis, and/or treatment of anemias and leukopeniasdescribed below. Alternatively, the modified transferrin fusion proteinsof the invention and/or polynucleotides encoding transferrin fusionproteins of the invention maybe used to decrease the quantity of all orsubsets of blood cells, such as, for example, erythrocytes, lymphocytes(B or T cells), myeloid cells (e.g., basophils, eosinophils,neutrophils, mast cells, macrophages) and platelets. The ability todecrease the quantity of blood cells or subsets of blood cells may beuseful in the prevention, detection, diagnosis, and/or treatment ofleukocytoses, such as, for example eosinophilia. The modified fusionproteins of the invention and/or polynucleotides encoding transferrinfusion proteins of the invention may be used to prevent, treat, ordiagnose blood dyscrasia.

Anemias are conditions in which the number of red blood cells or amountof hemoglobin (the protein that carries oxygen) in them is below normal.Anemia may be caused by excessive bleeding, decreased red blood cellproduction, or increased red blood cell destruction (hemolysis). Themodified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in treating, preventing, and/or diagnosing anemias.Anemias that may be treated prevented or diagnosed by the transferrinfusion proteins of the invention and/or polynucleotides encodingtransferrin fusion proteins of the invention include iron deficiencyanemia, hypochromic anemia, microcytic anemia, chlorosis, hereditarysideroblastic anemia, idiopathic acquired sideroblastic anemia, red cellaplasia, megaloblastic anemia (e.g., pernicious anemia, (vitamin B12deficiency) and folic acid deficiency anemia), aplastic anemia,hemolytic anemias (e.g., autoimmunune hemolytic anemia, microangiopathichemolytic anemia, and paroxysmal nocturnal hemoglobinuria). The modifiedtransferrin fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention may be useful intreating, preventing, and/or diagnosing anemias associated with diseasesincluding but not limited to, anemias associated with systemic lupuserythematosus, cancers, lymphomas, chronic renal disease, and enlargedspleen. The transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in treating, preventing, and/or diagnosing anemia arisingfrom drug treatments such as anemias associated with methyldopa,dapsone, and/or sulfa drugs. Additionally, modified fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention maybe useful in treating, preventing, and/ordiagnosing anemias associated with abnormal red blood cell architectureincluding but not limited to, hereditary spherocytosis, hereditaryelliptocytosis, glucose-6-phosphate dehydrogenase deficiency, and sicklecell anemia.

The modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in treating, preventing, and/or diagnosing hemoglobinabnormalities, (e.g., those associated with sickle cell anemia,hemoglobin C disease, hemoglobin S-C disease, and hemoglobin E disease).Additionally, the transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful in diagnosing, preventing, and/or prognosing in treatingthalassemias, including, but not limited to, major and minor forms ofalpha-thalassemia and beta-thalassemia.

In another embodiment, the modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be useful in diagnosing, prognosing, preventing,and/or treating bleeding disorders including, but not limited to,thrombocytopenia (e.g., idiopathic thrombocytopenic purpura, andthrombotic thrombocytopenic purpura), Von Willebrand's disease,hereditary platelet disorders (e.g., storage pool disease such asChediak-Higashi and Hermansky-Pudlak syndromes, thromboxane A2dysfunction, thromboasthenia, and Bernard-Soulier syndrome),hemolyticuremic syndrome, hemophilias such as hemophilia A or Factor V11deficiency and Christmas disease or Factor IX deficiency, HereditaryHemorhhagic Telangiectsia, also known as Rendu-Osler-Webe syndrome,allergic purpura (Henoch Schonlein purpura) and disseminatedintravascular coagulation.

In other embodiments, the modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be useful as an agent to increase cytokine production.

In certain embodiments, fusion proteins of the invention, and/orpolynucleotides encoding transferrin fusion proteins of the inventioncan be used to treat or detect hyperproliferative disorders, includingneoplasms. Transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay inhibit the proliferation of the disorder through direct or indirectinteractions. Alternatively, fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay cause proliferation of other cells which can inhibit thehyperproliferative disorder.

For example, by increasing an immune response, particularly increasingantigenic qualities of the hyperproliferative disorder or byproliferating, differentiating, or mobilizing T-cells,hyperproliferative disorders can be treated. This immune response may beincreased by either enhancing an existing immune response, or byinitiating a new immune response. Alternatively, decreasing an immuneresponse may also be a method of treating hyperproliferative disorders,such as a chemotherapeutic agent.

Examples of hyperproliferative disorders that can be treated or detectedby modified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention include, but arenot limited to neoplasms located in the colon, abdomen, bone, breast,digestive system, liver, pancreas, peritoneum, endocrine glands(adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid),eye, head and neck, nervous (central and peripheral), lymphatic system,pelvis, skin, soft tissue, spleen, thorax, and urogenital tract.

Similarly, other hyperproliferative disorders can also be treated ordetected by modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the invention.Examples of such hyperproliferative disorders include, but are notlimited to Acute Childhood Lymphoblastic Leukemia; Acute LymphoblasticLeukemia, Acute Lymphocytic Leukemia, Acute Myeloid Leukemia,Adrenocortical Carcinoma, Adult (Primary) Hepatocellular Cancer, Adult(Primary) Liver Cancer, Adult Acute Lymphocytic Leukemia, Adult AcuteMyeloid Leukemia, Adult Hodgkin's Disease, Adult Hodgkin's Lymphoma,Adult Lymphocytic Leukemia, Adult Non-Hodgkin's Lymphoma, Adult PrimaryLiver Cancer, Adult Soft Tissue Sarcoma, AIDS-Related Lymphoma,AIDS-Related Malignancies, Anal Cancer, Astrocytoma, Bile Duct Cancer,Bladder Cancer, Bone Cancer, Brain Stem Glioma, Brain Tumors, BreastCancer, Cancer of the Renal Pelvis and Ureter, Central Nervous System(Primary) Lymphoma, Central Nervous System Lymphoma, CerebellarAstrocytoma, Cerebral Astrocytoma, Cervical Cancer, Childhood (Primary)Hepatocellular Cancer, Childhood (Primary) Liver Cancer, Childhood AcuteLymphoblastic Leukemia, Childhood Acute Myeloid Leukemia, ChildhoodBrain Stem Glioma, Childhood Cerebellar Astrocytoma, Childhood CerebralAstrocytoma, Childhood Extracranial Germ Cell Tumors, ChildhoodHodgkin's Disease, Childhood Hodgkin's Lymphoma, Childhood Hypothalamicand Visual Pathway Glioma, Childhood Lymphoblastic Leukemia, ChildhoodMedulloblastoma, Childhood Non-Hodgkin's Lymphoma, Childhood Pineal andSupratentorial Primitive Neuroectodermal Tumors, Childhood Primary LiverCancer, Childhood Rhabdomyosarcoma, Childhood Soft Tissue Sarcoma,Childhood Visual Pathway and Hypothalamic Glioma, Chronic LymphocyticLeukemia, Chronic Myelogenous Leukemia, Colon Cancer, Cutaneous T-CellLymphoma, Endocrine Pancreas Islet Cell Carcinoma. Endometrial Cancer,Ependymoma, Epithelial Cancer, Esophageal Cancer, Ewing's Sarcoma andRelated Tumors, Exocrine Pancreatic Cancer, Extracranial Germ CellTumor, Extragonadal Germ Cell Tumor, Extrahepatic Bile Duct Cancer, EyeCancer, Female Breast Cancer, Gaucher's Disease, Gallbladder Cancer,Gastric Cancer, Gastrointestinal Carcinoid Tumor, GastrointestinalTumors, Germ Cell Tumors, Gestational Trophoblastic Tumor, Hairy CellLeukemia, Head and Neck Cancer, Hepatocellular Cancer, Hodgkin'sDisease, Hodgkin's Lymphoma, Hypergammaglobulinemia, HypopharyngealCancer, Intestinal Cancers, Intraocular Melanoma, Islet Cell Carcinoma,Islet Cell Pancreatic Cancer, Kaposi's Sarcoma, Kidney Cancer, LaryngealCancer, Lip and Oral Cavity Cancer, Liver Cancer, Lung Cancer, Lymphoproliferative Disorders, Macroglobulinemia, Male Breast Cancer,Malignant Mesothelioma, Malignant Thymoma, Medulloblastomia, Melanoma,Mesothelioma, Metastatic Occult Primary Squamous Neck Cancer, MetastaticPrimary Squamous Neck Cancer, Metastatic Squamous Neck Cancer, MultipleMyeloma, Multiple Myeloma/Plasma Cell Neoplasm, MyelodysplasticSyndrome, Myelogenous Leukemia, Myeloid Leukemia, MyeloproliferativeDisorders, Nasal Cavity and Paranasal Sinus Cancer, NasopharyngealCancer, Neuroblastoma, Non-Hodgkin's Lymphoma During Pregnancy,Nonmelanoma Skin Cancer, Non-Small Cell Lung Cancer, Occult PrimaryMetastatic Squamous Neck Cancer, Oropharyngeal Cancer, Osteo/MalignantFibrous Sarcoma, Osteosarcoma/Malignant Fibrous Histiocytoma,Osteosarcoma/Malignant Fibrous Histiocytoma of Bone, Ovarian EpithelialCancer, Ovarian Germ Cell Tumor, Ovarian Low Malignant Potential Tumor,Pancreatic Cancer, Paraproteinemias, Purpura, Parathyroid, Cancer,Penile Cancer, Pheochromocytoma, Pituitary Tumor, Plasma CellNeoplasm/Multiple Myeloma, Primary Central Nervous System Lymphoma,Primary Liver Cancer, Prostate Cancer, Rectal Cancer, Renal Cell Cancer,Renal Pelvis and Ureter Cancer, Retinoblastoma, Rhabdomyosarcoma,Salivary Gland Cancer, Sarcoidosis Sarcomas, Sezary Syndrome, SkinCancer, Small Cell Lung Cancer, Small Intestine Cancer, Soft TissueSarcoma, Squamous Neck Cancer, Stomach Cancer, Supratentorial PrimitiveNeuroectodermal and Pineal Tumors, T-Cell Lymphoma, Testicular Cancer,Thymoma, Thyroid Cancer, Transitional Cell Cancer of the Renal Pelvisand Ureter, Transitional Renal Pelvis and Ureter Cancer, TrophoblasticTumors, Ureter and Renal Pelvis Cell Cancer, Urethral Cancer, UterineCancer, Uterine Sarcoma, Vaginal Cancer, Visual Pathway and HypothalamicGlioma, Vulvar Cancer, Waldenstrom's Macroglobulinemia, Wilm's Tumor,and any other hyperproliferative disease, besides neoplasia, located inan organ system listed above.

In another preferred embodiment, modified transferrin fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention are used to diagnose, prognose, prevent,and/or treat premalignant conditions and to prevent progression to aneoplastic or malignant state, including but not limited to thosedisorders described above. Such uses are indicated in conditions knownor suspected of preceding progression to neoplasia or cancer, inparticular, where non-neoplastic cell growth is consisting ofhyperplasia, metaplasia, or most particularly, dysplasia has occurred(for review of such abnormal growth conditions, see Robbins. and Angell,1976, Basic Pathology, 2d Ed. W. B. Saunders Co., Philadelphia, pp.68-79).

Hyperplasia is a form of controlled cell proliferation, involving anincrease in cell number in a tissue or organ, without significantalteration in structure or function. Hyperplastic disorders which can bediagnosed, prognosed, prevented, and/or treated with fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention include, but are not limited to,angiofollicular mediastinal lymph node hyperplasia, angiolymphoidhyperplasia with eosinophilia, atypical melanocytic hyperplasia, basalcell hyperplasia, benign giant lymph node hyperplasia, cementumhyperplasia, congenital adrenal hyperplasia, congenital sebaceoushyperplasia, cystic hyperplasia, cystic hyperplasia of the breast,denture hyperplasia, ductal hyperplasia, endometrial hyperplasia,fibromuscular hyperplasia, foca epithelial hyperplasia, gingivalhyperplasia, inflammatory fibrous hyperplasia, inflammatory papillaryhyperplasia, intravascular papillary endothelial hyperplasia, nodularhyperplasia of prostate, nodular regenerative hyperplasia,pseudoepitheliomatous hyperplasia, senile sebaceous hyperplasia, andverrucous hyperplasia.

In another embodiment, modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention conjugated to a toxin or a radio-active isotope, asdescribed herein, may be used to treat cancers and neoplasms, including,but not limited to, those described herein. In a further preferredembodiment, transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionconjugated to a toxin or a radioactive isotope, as described herein, maybe used to treat acute myelogenous leukemia.

Additionally, modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay affect apoptosis, and therefore, would be useful in treating anumber of diseases associated with increased cell survival or theinhibition of apoptosis. For example, diseases associated with increasedcell survival or the inhibition of apoptosis that could be diagnosed,prognosed, prevented, and/or treated by polynucleotides, polypeptides,and/or agonists or antagonists of the invention, include cancers (suchas follicular-lymphomas, carcinomas with p53 mutations, andhormone-dependent tumors, including, but not limited to, colon cancer,cardiac tumors, pancreatic cancer, melanoma, retinoblastoma,glioblastoma, lung cancer, intestinal cancer, testicular cancer, stomachcancer, neuroblastoma, myxoma, myoma, lymphoma, endothelioma,osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, adenoma,breast cancer, prostrate cancer, Kaposi's sarcoma and ovarian cancer);autoimmune disorders such as, multiple sclerosis, Sjogren's syndrome,Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn'sdisease, polymyositis, systemic lupus erythematosus and immune-relatedglomerulonephritis and rheumatoid arthritis) and viral infections (suchas herpes viruses, pox viruses and adenoviruses), inflammation, graft v.host disease, acute graft rejection, and chronic graft rejection.

In preferred embodiments, modified fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention are used to inhibit growth, progression, and/or metastasis ofcancers, in particular those listed above.

Additional diseases or conditions associated with increased cellsurvival that could be diagnosed, prognosed, prevented, and/or treatedby modified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention, include but arenot limited to, progression and/or metastases of malignancies andrelated disorders such as leukemia (including acute leukemia (e.g.,acute lymphocytic leukemia, acute myelocytic leukemia (includingmyeloblastic, promyelocytic, mylomonocytic, monocytic, anderythroleukemia)) and chronic leukemia (e.g., chronic myelocytic(granulocytic) leukemia and chronic lymphocytic leukemia)), polycythemiavera, lymphomas (e.g., Hodgkin's disease and non-Hodgkin's disease),multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease,and solid tumors including, but not limited to, sarcomas and carcinomassuch as fibrosarcoma, myxosarcoma, fiposarcoma, chondrosarcoma,osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma,lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma,Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma,pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, papillary carcinoma,papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma,bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile ductcarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor,cervical cancer, testicular tumor, lung carcinoma, small cell lungcarcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,medulloblastoma, craniopharyngioma, ependymoma, pinealoma,emangioblastoma, acoustic neuroma, oligodendrogliomia, menangioma,melanoma, neuroblastoma, and retinoblastoma.

Diseases associated with increased apoptosis that could be diagnosed,prognosed, prevented, and/or treated by modified fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention, include AIDS; neurodegenerative disorders (such asAlzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis,retinitis pigmentosa, cerebral degeneration and brain tumor or prionassociated disease); autoimmune disorders (such as, multiple sclerosis,Sjogren's syndrome, Hashimoto's thyroiditis, biliary cirrhosis, Behcet'sdisease, Crohn's disease, polymyositis, systemic lupus erythematosus andimmune-related glomerulonephritis and rheumatoid arthritis)myelodysplastic syndromes (such as aplastic anemia), graft Y hostdisease, ischemic injury (such as that caused by myocardial infarction,stroke and repercussion injury), liver injury (e.g., hepatitis relatedliver injury, ischemia/reperfusion injury, cholestosis (bile ductinjury) and liver cancer); toxin-induced liver disease (such as thatcaused by alcohol), septic shock, cachexia and anorexia.

Another preferred embodiment utilizes polynucleotides encoding modifiedtransferrin fusion proteins of the invention to inhibit aberrantcellular division, by gene therapy using the present invention, and/orprotein fusions or fragments thereof.

Thus, the present invention provides a method for treating cellproliferative disorders by inserting into an abnormally proliferatingcell a polynucleotide encoding modified transferrin fusion protein ofthe present invention, wherein said polynucleotide represses saidexpression.

Another embodiment of the present invention provides a method oftreating cell proliferative disorders in individuals comprisingadministration of one or more active gene copies of the presentinvention to an abnormally proliferating cell or cells.

The polynucleotides of the present invention may be delivered directlyto cell proliferative disorderly disease sites in internal organs, bodycavities, and the like by use of imaging devices used to guide aninjecting needle directly to the disease site. The polynucleotides ofthe present invention may also be administered to disease sites at thetime of surgical intervention.

By cell proliferative disease is meant any human or animal disease ordisorder, affecting any one or any combination of organs, cavities, orbody parts, which is characterized by single or multiple local abnormalproliferations of cells, groups of cells, or tissues, whether benign ormalignant.

Any amount of the polynucleotides of the present invention may beadministered as long as it has a biologically inhibiting effect on theproliferation of the treated cells.

Moreover, it is possible to administer more than one of thepolynucleotides of the present invention simultaneously to the samesite. By “biologically inhibiting” is meant partial or total growthinhibition as well as decreases in the rate of proliferation or growthof the cells.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionare useful in inhibiting the metastasis of proliferative cells ortissues. Inhibition may occur as a direct result of administering thesetransferrin fusion proteins and/or polynucleotides, or indirectly, suchas activating the expression of proteins known to inhibit metastasis,for example alpha, integrins, (See, e.g., Curr. Top. Mirobiol. Immunol.1998; 231:1 41, which is hereby incorporated by reference). Suchtherapeutic affects of the present invention may be achieved eitheralone, or in combination with small molecule drugs or adjuvants.

In another embodiment, the invention provides a method of deliveringcompositions containing the transferrin fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention to targeted cells expressing a polypeptide bound by, thatbinds to, or associates with a modified transferrin fusion protein ofthe invention. Transferrin fusion proteins of the invention may beassociated with heterologous polypeptides, heterologous nucleic acids,toxins, or prodrugs via hydrophobic, hydrophilic, ionic and/or covalentinteractions.

Kidney diseases which can be diagnosed, prognosed, prevented, and/ortreated with compositions of the invention include, but are not limitedto, acute kidney failure, chronic kidney failure, atheroembolic renalfailure, end-stage renal disease, inflammatory diseases of the kidney(e.g., acute glomerulonephritis, post infectious glomerulonephritis,rapidly progressive glomerulonephritis, nephritic syndrome, membranousglomerulonephritis, familial nephritic syndrome, membrane proliferativeglomerulonephritis and mesangial proliferative glomerulonephritis,chronic glomerulonephritis, acute tubulo-interstitial nephritis, chronictubulointerstitial nephritis, acute post-streptococcalglomerulonephritis (PSGN), pyelonephritis, lupus nephritis, chronicnephritis, interstitial nephritis, and post streptococcalglomerulonephritis), blood vessel disorders of the kidneys (e.g., kidneyinfarction, atheroembolic kidney disease, cortical necrosis, malignantnephrosclerosis, renal vein thrombosis, renal under perfusion, renalretinopathy, renal ischemia-reperfusion, renal artery embolism and renalartery stenosis), and kidney disorders resulting form urinary tractdisease (e.g., pyelonephritis, hydronephrosis, urolithiasis (renallithiasis, nephrolithiasis), reflux nephropathy, urinary tractinfections, urinary retention, and acute or chronic unilateralobstructive uropathy). In addition, compositions of the invention can beused to diagnose, prognose, prevent, and/or treat metabolic andcongenital disorders of the kidney (e.g., uremia, renal amyloidosis,renal osteodystrophy, renal tubular acidosis, renal glycosuria,nephrogenic diabetes insipidus, cystinuria, Fanconi's syndrome, renalfibrocystic osteosis (renal rickets), Hartnup disease, Bartter'ssyndrome, Liddle's syndrome, polycystic kidney disease, medullary cysticdisease, medullary sponge kidney, Alport's syndrome, nail-patellasyndrome, congenital nephritic syndrome, CRUSH syndrome, horseshoekidney, diabetic nephropathy, nephrogenic diabetes insipidus, analgesicnephropathy, kidney stones, and membranous nephropathy), and autoimmunedisorders of the kidney (e.g., systemic lupus erythematosus (SLE), Goodpasture syndrome, IgA nephropathy, and ICFM mesangial proliferativeglomerulonephritis).

Compositions of the invention can also be used to diagnose, prognose,prevent, and/or treat sclerotic or lecrotic disorders of the kidney(e.g., glomerulosclerosis, diabetic nephropathy, focal Fsegmentalglomerulo sclerosis (FSGS), narcotizing glomerulonephritis, and renalpapillary necrosis), cancers of the kidney (e.g., nephroma,hypernephroma, nephroblastoma, renal cell cancer, transitional cellcancer, renal adenocarcinoma, squamous cell cancer, and Wilm's tumor),and electrolyte imbalances (e.g., nephrocalcinosis, pyuria, edema,hydronephritis, proteinuria, hyponatremia, hypernatremia, hypokalemia,hyperkalemia, hypocalcemia, hypercalcemia, hypophosphatemia, andhyperphosphatemia).

Compositions of the invention may be administered using any method knownin the art, including, but not limited to, direct needle injection atthe delivery site, intravenous injection, topical administration,catheter infusion, biolistic injectors, particle accelerators, gel foamsponge depots, other commercially available depot materials, osmoticpumps, oral or suppositorial solid pharmaceutical formulations,decanting or topical applications during surgery, aerosol delivery. Suchmethods are known in the art. Compositions of the invention may beadministered as part of a Therapeutic, described in more detail below.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the invention,may be used to treat, prevent, diagnose, and/or prognose cardiovasculardisorders, including, but not limited to, peripheral artery disease,such as limb ischemia.

Cardiovascular disorders, includes, but is not limited to,cardiovascular abnormalities, such as arterio arterial fistula,arteriovenous fistula, cerebral arteriovenous malformations, congenitalheart defects, pulmonary atresia, and Scimitar Syndrome.

Congenital heart defects include, but are not limited to, aorticcoarctation, cortriatriatum, coronary vessel anomalies, crisscrossheart, dextrocardia, patent ductus arteriosus, Ebstein's anomaly,Eisenmenger complex, hypoplastic left heart syndrome, levocardia,tetralogy of fallot, transposition of great vessels, double outlet rightventricle, tricuspidatresia, persistent truncus arteriosus, and heartseptal defects, such as aortopulmonary septald defect, endocardialcushion defects, Lutembacher's Syndrome, trilogy of Fallot, ventricularheart septal defects.

Cardiovascular disorders also include, but are not limited to, heartdisease, such arrhythmias, carcinoid heart disease, high cardiac output,low cardiac output, cardiactamponade, endocarditis (including bacteria),heart aneurysm, cardiac arrest, congestive heart failure, congestivecardiomyopathy, paroxysmal dyspnea, cardiac edema, heart hypertrophy,congestive cardiomyopathy left ventricular hypertrophy, rightventricularhypertrophy, post-infarction heart rupture, ventricularseptal rupture, heart valve diseases myocardial diseases, myocardialischemia, pericardial effusion, pericarditis (including constrictive andtuberculous), pricumopericardium, post pericardiotomy syndrome,pulmonary heart disease, rheumatic heart disease, ventriculardysfunction, hyperemia, cardiovascular pregnancy complications, ScimitarSyndrome, cardiovascular syphilis, and cardiovascular tuberculosis.

Arrhythmias include, but are not limited to, sinus arrhythmia, atrialfibrillation, atrial flutter, bradycardia, extrasystole, Adams-StokesSyndrome, bundle-branch block, sinoatrial block, long QT syndrome,parasystole, Lown-Ganong-Levine Syndrome, Mahaim-type pre-excitationsyndrome, Wolff-Parkinson-White syndrome, sick sinus syndrome,itachycardias, and ventricular fibrillation. Tachycardias includeparoxysmal tachycardia, suprayentriculai tachycardia, acceleratedidioventricular rhythm, atrioventricular nodal reentry tachyeardia,ectopic atrial tachycardia, ectopic junctional tachycardia, sinoattialnodalreentry tachycardia, sinus tachycardia, Torsades de Pointes, andventricular tachycardia.

Heart valve diseases include, but are not limited to, aortic valveinsufficiency aortic valve stenosis, heart murmurs, aortic valveprolapse, neutral valve prolapse, tricuspid valve prolapse, mitral valveinsufficiency, mitral valve stenosis, pulmonary atresia, pulmonary valveinsufficiency, pulmonary valve stenosis, tricuspid atresia, tricuspidvalve insufficiency, and tricuspid valve stenosis.

Myocardial diseases include, but are not limited to, alcoholiccardiomyopathy, congestive cardiomyopathy, hypertrophic cardiomyopathy,aortic subvalvular stenosis, pulmonary subvalvular stenosis, restrictivecardiomyopathy, Chagas cardiomyopathy, endocardial fibroelastosis,endomyocardial fibrosis, Kearns Syndrome, myocardial reperfusion injury,and myocarditis.

Myocardial ischemias include, but are not limited to, coronary disease,such as angina pectoris, coronary aneurysm, coronary arteriosclerosis,coronary thrombosis, coronary vasospasm, myocardial infarction, andmyocardial stunning.

Cardiovascular diseases also include vascular diseases such asaneurysms, angiodysplasia, angiomatosis, bacillary angiomiatosis,Hippel-Lindau Disease, Klippel Trenaunay Weber Syndrofne, Sturge WeberSyndrome, angioneurotic edema, aortic diseases, Takayasu's Arthritis,aortitis, Leriche's Syndrome, arterial occlusive diseases, arthritis,enarteritis, polyarteritis nodosa, cerebrovascular disorders, diabeticangiopathies, diabetic retinopathy, embolisms, thrombosis,erythromelalgia, hemorrhoids, hepatic veno-occlusive disease,hypertension, hypotension, ischemia, peripheral vascular diseases,phlebitis, pulmonary veno-occlusive disease, Raynaud's disease, CRESTsyndrome, retinal vein occlusion, Scimitar syndrome, superior vena cavasyndrome, telangiectasia, ataxia telangiectasia, hereditary hemorrhagictelangiectasia, varicocele, varicose veins, varicoseulcer, vasculitis,and venous insufficiency.

Cerebrovascular disorders include, but are not limited to, cardio arterydiseases but includes respiratory disorders. Transferrin fusion proteinsof the invention and/or polynucleotides encoding transferrin fusionproteins of the invention may be used to treat, prevent, diagnose,and/or prognose diseases and/or disorders of the respiratory system.

Diseases and disorders of the respiratory system include, but are notlimited to, nasalvestibulitis, nonallergic rhinitis (e.g., acuterhinitis, chronic rhinitis, atrophic rhinitis, vasomotor rhinitis),nasal polyps, and sinusitis, juvenile angiofibromas, cancer of the noseand juvenile papillomas, vocal cord polyps, nodules (singer's nodules),contact ulcers, vocal cord paralysis, laryngoceles, pharyngitis (e.g.,viral and bacterial), tonsillitis, tonsillar cellulitis, parapharyngealabscess, laryngitis, laryngoceles, and throat cancers (e.g., cancer ofthe nasopharynx, tonsil cancer, larynx cancer), lung cancer (e.g.,squamous cell carcinoma, small cell (oat cell) carcinoma, large cellcarcinoma, and adenocarcinoma), allergic disorders (eosinophilicpneumonia, hypersensitivity pneumonitis (e.g., extrinsicallergicalveolitis, allergic interstitial pneumonitis, organic dustpneumoconiosis, allergic bronchopulmoniary aspergillosis, asthma,Wegener's granulomatosis (granulomatous vasculitis), Goodpasture'ssyndrome), pneumonia (e.g., bacterial pneumonia (e.g., Streptococcuspneumoniae (pneumoncoccal pneumonia), Staphylococcus aureus(staphylococeal pneumonia), Gram negative bacteria pneumonia (caused by,e.g., Klebsiella and Pseudomonas spp.), Mycoplasma pneumoniae pneumonia,Hemophilus influenza pneumonia, Legionella pneumophila (Legionnaires'disease), and Chlamydia psittaci (Psittacosis)), and viral pneumonia(e.g., influenza, chickenpox (varicella).

Additional diseases and disorders of the respiratory system include, butare not limited to bronchiolitis, polio (poliomyelitis), croup,respiratory syncytial viral infection, mumps, erythema infectiosum(fifth disease), roseola infantum, progressive rubellapanencephalitis,German measles, and subacute sclerosing panencephalitis), fungalpneumonia (e.g., Histoplasmosis, Coccidioidomycosis, Blastomycosis,fungal infections in people with severely suppressed immune systems(e.g., cryptococcosis, caused by Cryptococcus neoformans; aspergillosis,caused by Aspergillus spp.) candidiasis, caused by Candida; andmucormycosis)), Pneumocystis carinii (pneumocystis pneumonia),atypicalpneumonias (e.g., Mycoplasma and Chlamydia spp.), opportunisticinfection pneumonia, nosocomial pneumonia, chemical pneumonitis, andaspiration pneumonia, pleural disorders (e.g., pleurisy, pleuraleffusion, and pneumothorax (e.g., simple spontaneous pneumothorax,complicated spontaneous pneumothorax, tension pneumothorax)),obstructive airway diseases (e.g., asthma, chronic obstructive pulmonarydisease (COPID), emphysema, chronic or acute bronchitis), occupationallung diseases (e.g., silicosis, blacklung (coal workers' pneumoconiosis,asbestosis, berylliosis, occupational asthma, and byssinosis),Infiltrative Lung Disease (e.g., pulmonary-fibrosis (e.g., usualinterstitial pneumonia), idiopathic pulmonary fibrosis, desquamativeinterstitial pneumonia, lymphoid interstitial pneumonia, histiocytosis(e.g., Letterer-Siwe disease, Hand-Schüller-Christian disease,eosinophilic granuloma), idiopathic pulmonary hemosiderosis, sarcoidosisand pulmonary, alveolar proteinosis), Acute respiratory distresssyndrome (also called, e.g., adult respiratory distress syndrome),edema, pulmonary embolism, bronchitis (e.g., viral, bacterial),bronchiectasis, atelectasis, lung abscess (caused by, e.g.,Staphylococcus aureus or Legionella pneumophila), and cystic fibrosis.

Cancers which may be treated with modified fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention include, but are not limited to solid tumors, includingprostate, lung, breast, ovarian, stomach, pancreas, larynx, esophagus,liver, parotid, biliary tract, colon, rectum, cervix, uterus,endometrium, kidney, bladder, thyroid cancer; primary tumors andmetastases; melanomas; glioblastoma; Kaposi's sarcoma; leiomyosarcoma;non-small cell lung cancer; colorectal cancer; advanced malignancies;and blood born tumors such as leukemia. For example, fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention may be delivered topically, in order to treatcancers such as skin cancer, head and neck tumors, breast tumors, andKaposi's sarcoma.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be useful, in treating other disorders, besides cancers, whichinvolve angiogenesis. These disorders include, but are not limited to:benign tumors, for example hemangiomas, acoustic neuromas,neurofibromas, trachomas, and pyogenicgranulomas; artherosclericplaques; ocular angiogenic diseases, for example, diabetic retinopathy,retinopathy of prematurity, macular degeneration, corneal graftrejection, neovascular glaucoma, retrolental fibroplasia, rubeosis,retinoblastoma, uvietis and Pterygiaab normal blood vessel growth) ofthe eye; rheumatoid arthritis; psoriasis; delayed wound healing;endometriosis; vasculogenesis; granulations; hypertrophic scars(keloids); nonunion fractures; scleroderma; trachoma; vascularadhesions; myocardial angiogenesis; coronary collaterals; cerebralcollaterals; arteriovenous malformations; ischemic limb angiogenesis;Osler-Webber Syndrome; plaque neovascularization; telangiectasia;hemophiliac joints; angiofibroima; fibromuscular dysplasia; woundgranulation; Crohn's disease; and atherosclerosis.

Thus, within one aspect of the present invention methods are providedfor treating neovascular diseases of the eye.

Additionally, disorders which can be treated with modified fusionproteins of the invention and/or polynucleotides encoding transferrinfusion proteins of the invention include, but are not limited to,hemangioma, arthritis, psoriasis, angiofibroma, atherosclerotic plaques,delayed wound healing, granulations, hemophilic joints hypertrophicscars, nonunion fractures, Osler-Weber syndrome, pyogenic granuloma,scleroderma, trachoma; and vascular adhesions.

Moreover, disorders and/or states, which can be treated, prevented,diagnosed, and/or prognosed with the modified transferrin fusionproteins of the invention and/or polynucleotides encoding transferrinfusion proteins of the invention include, but are not limited to, solidtumors, blood born tumors such as leukemia, tumor metastasis, Kaposi'ssarcoma, benign tumors, for example hemangiomas, acoustic neuromas,neurofibromas, trachomas, and pyogenic granulomas, rheumatoid arthritis,psoriasis, ocularangiogenic diseases, for example, diabetic retinopathy,retinopathy of prematurity, macular degeneration, corneal graftrejection, neovascular glaucoma, retrolental fibroplasia, rubeosis,refinoblastoma, and uvietis, delayed wound healing, endometriosis,vasculogenesis, granulations, hypertrophic scars (keloids, nonunionfractures, sclerodemm, trachoma, vascular adhesions, myocardialangiogenesis, coronary collaterals, cerebral collaterals, arteriovenousmalformations, ischemic limb angiogenesis, Osler-Webber Syndrome, plaqueneovascularization, telangiectasia, hemophiliac joints, angiofibromafibromuscular dysplasia, wound granulation, Crohn's disease,atherosclerosis, birth control agent by preventing vascularizationrequired for embryo, implantation controlling menstruation, diseasesthat have angiogenesis as a pathologic consequence such as cat scratchdisease (Rochele nunalia quintosa), ulcers (Helicobacter pylori),Bartonellosis and baculary angiomatosis.

In one aspect of the birth control method, an amount of the compoundsufficient to block embryo implantation is administered before or afterintercourse and fertilization have occurred, thus providing an effectivemethod of birth control, possibly a “morning after” method. Modifiedtransferrin fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention may also be usedin controlling menstruation or administered as either a peritoneallavage fluid or for peritoneal implantation in the treatment ofendometriosis.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be utilized in a wide variety of surgical procedures.

Diseases associated with increased cell survival or the inhibition ofapoptosis that could be treated, prevented, diagnosed, and/or prognosedusing modified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention, include cancers(such as follicular lymphomas, carcinomas with mutations, andhormone-dependent tumors, including, but not limited to colon cancer,cardiac tumors, pancreatic cancer, melanoma, retinoblastoma,glioblastoma, lung cancer, intestinal cancer, testicular cancer, stomachcancer, neuroblastoma, myxoma, myoma, lymphoma, endothelioma,osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, adenoma,breast cancer, prostate cancer, Kaposi's sarcoma and ovarian cancer);autoimmune disorders (such as, multiple sclerosis, Sjogren's syndrome,Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn'sdisease, polymyositis, systemic lupus thematosus and immune-relatedryglomerulonephritis and rheumatoid arthritis) and viral infections(such as herpes viruses, pox viruses and adenoviruses), inflammation,graft v. host disease, acute graft rejection, and chronic graftrejection.

In preferred embodiments, modified fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention are used to inhibit growth, progression, and/or metasis ofcancers, in particular those listed above.

Additional diseases or conditions associated with increased cellsurvival that could be treated or detected by modified fusion proteinsof the invention and/or polynucleotides encoding, transferrin fusionproteins of the invention include, but are not limited to, progression,and/or metastases of malignances and related disorders such as leukemia(including acute leukemia (e.g., acute lymphocytic leukemia, acutemyelocytic leukemia (including myeloblastic, promyelocytic,myelomonocytic, monocytic, and erythroleukemia)) and chronic leukemia(e.g., chronic myelocytic (granulocytic) leukemia and chroniclymphocyticleukemia)), polycytemia vera, lymphomas (e.g., Hodgkin's disease andnon-Hodgkin's disease), multiple myeloma, Waldenstrom'smacroglobulinemia, heavy chain disease, and solid tumors including, butnot limited to, Sarcomas and carcinomas such as fibrosarcoma,myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma,angiosarcoma, endotheliosarcoma, lymphangiosarcoma,lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor,leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer,breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma,basa cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceousgland carcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct Carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testiculartumor, Jung carcinoma, small cell lung carcinoma, bladder carcinoma,epithelial carcinoma, glioma, astrocytoma, medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, menangioma, melanoma neuroblastoma, andretinoblastoma.

Diseases associated with increased apoptosis that could be treated,prevented, diagnosed, and/or prognosed using modified fusion proteins ofthe invention and/or polynucleotides encoding transferrin fusionproteins of the invention, include, but are not limited to, AIDS;neurodegenerative disorders (such as Alzheimer's disease, Parkinson'sdisease, Amyotrophic lateral sclerosis, Retinitis pigmentosa, Cerebellardegeneration and brain tumor or prion associated disease); autoimmunedisorders (such as, multiple sclerosis, Sjogren's syndrome, Hashimoto'sthyroiditis, biliary cirrhosis, Behcet's disease, Crohn's′ disease,polymyositis, systemic lupus erythematosus and immune-relatedglomerulonephritis and rheumatoid arthritis) Myelodysplastic syndromes(such as aplastic anemia), graft v. host disease, ischemic injury (suchas that caused, by myocardial infarction, stroke and reperfusioninjury), liver injury (e.g., hepatitis related liver injury,ischemia/reperfusion injury, cholestosis (bile duct injury) and livercancer); toxin-induced liver disease (such as that caused by alcohol),septic shock, cachexia and anorexia.

In addition, modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventioncould be, used to treat or prevent the onset of diabetes mellitus. Inpatients with newly diagnosed Types I and II diabetes, where some isletcell function remains, fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the invention,could be used to maintain the islet function so as to alleviate, delayor prevent permanent manifestation of the disease. Also, fusion proteinsof the invention and/or polynucleotides encoding transferrin fusionproteins of the invention could be used as an auxiliary in islet celltransplantation to improve or promote islet cell function.

The modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be used for the diagnosis and/or treatment of diseases, disorders,damage or injury of the brain and/or nervous system. Nervous systemdisorders that can be treated with the compositions of the invention(e.g., fusion proteins of the invention and/or polynucleotides encodingtransferrin fusion proteins of the invention), limited to nervoussystems include, but are not limited injuries, and diseases or disorderswhich result in either a disconnection of axons, a diminution ordegeneration of neurons, or demyelination. Nervous system lesions whichmay be treated in a patient (including human and non-human mammalianpatients) according to the methods of the invention, include but are notlimited to, the following lesions of either the central (includingspinal cord, brain) or peripheral nervous systems: (1) ischemic lesions,in which a lack of oxygen in a portion of the nervous system results inneuronal injury or death, including cerebral infarction or ischemia, orspinal cord infarction or ischemia; (2) traumatic lesions, includinglesions caused by physical injury or associated with surgery, forexample, lesions which sever a portion of the nervous system, orcompression injuries; (3) malignant lesions, in which a portion of thenervous system is destroyed or injured by malignant tissue which iseither a nervous system associated malignancy or a malignancy derivedfrom nervous system tissue; (4) infectious lesions in which a portion ofthe nervous system is destroyed or injured as a result of infection, forexample, by an abscess or associated with infection by humanimmunodeficiency virus, herpes zoster, or herpes simplex virus or with,Lyme disease, tuberculosis, or syphilis; (5) degenerative lesions, inwhich a portion of the nervous system is destroyed or injured as aresult of a degenerative process including but not limited to,degeneration associated with Parkinson's disease, Alzheimer's disease,Huntington's chorea, or amyotrophic lateral sclerosis (ALS); (6) lesionsassociated with nutritional diseases or disorders, in which a portion ofthe nervous system is destroyed or injured by a nutritional disorder ordisorder of metabolism including, but not limited to vitamin B12deficiency, folic acid deficiency, Wernicke disease, tobacco-alcoholamblyopic, Marchiafava-Blanami disease (primary degeneration of thecorpus callosum), and alcoholic cerebral degeneration; (7) neurologicallesions associated with systemic diseases including, but not limited todiabetes (diabetic neuropathy, Bell's palsy), systemic lupuserythematosus, carcinoma, or sarcoidoisis; (8) lesions caused by toxicsubstances including alcohol, lead, or particular, neurotoxins; and (9)demyelinated lesions in which a portion of the nervous system isdestroyed or injured by a demyelinating disease including, but notlimited to, multiple sclerosis, human immunodeficiency virus-associatedmyelopathy, transverse myelopathy or various etiologies, progressivemultifocal leukoencephalopathy, and central pontine myelinolysis.

In one embodiment, the modified transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention are used to protect neural cells from the damaging effectsof hypoxia. In a further preferred embodiment, the modified transferrinfusion proteins of the invention and/or polynucleotides encodingtransferrin fusion proteins of the invention are used to protect neuralcells from the damaging effects of cerebral hypoxia.

In specific embodiments, motor neuron disorders that may be treatedaccording to the invention include, but are not limited to, disorderssuch as infarction, infection, exposure to toxin, trauma, surgicaldamage, degenerative disease or malignancy that may affect motor neuronsas well as other components of the nervous system, as well as disordersthat selectively affect neurons such as amyotrophic lateral sclerosis,and including, but not limited to, progressive spinal muscular atrophy,progressive bulbar palsy, primary lateral sclerosis, infantile andjuvenile muscular atrophy, progressive bulbar paralysis of childhood(Fazio-Londe syndrome), poliomyelitis and the post polio syndrome, andHereditary Motor sensory Neuropathy (Charcot-Marie-Tooth Disease).

Further, modified fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay play a role in neuronal survival; synapse formation; conductance;neural differentiation, etc. Thus, compositions of the invention(including fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention) may be used todiagnose and/or treat or prevent diseases or disorders associated withthese roles, including, but not limited to, learning and/or cognitiondisorders. The compositions of the invention may also be useful in thetreatment or prevention of neurodegenerative disease states and/orbehavioral disorders. Such neurodegenerative disease states and/orbehavioral disorders include, but are not limited to, Alzheimer'sDisease, Parkinson's Disease, Huntington's Disease, Tourette Syndrome,schizophrenia, mania, dementia, paranoia, obsessive compulsive disorder,panic disorder, learning disabilities, ALS, psychoses, autism, andaltered behaviors, including disorders in feeding, sleep patterns,balance, and perception.

Examples of neurologic diseases which can be treated or detected withmodified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention include, braindiseases, such as metabolic brain diseases which includesphenylketonuria such as maternal phenylketonuria, pyruvate carboxylasedeficiency, pyruvate dehydrogenase complex deficiency, Wernicke'sEncephalopathy, brain edema, brain neoplasms such as cerebellarneoplasms which include infratentorial neoplasms, cerebral ventricleneoplasms such as choroid plexus neoplasms, hypothalamic neoplasms,supratentorial neoplasms, canavan disease, cerebellar diseases such ascerebellar ataxia which include spinocerebellar degeneration such asataxia telangiectasia, cerebellar dyssynergia, Friederich's Ataxia,Machado-Joseph Disease, olivopontocerebellar atrophy, cerebellarneoplasms such as infratentorial neoplasms, diffuse cerebral sclerosissuch as encephalitis periaxialis, globoid cell leukodystrophy,metachromatic leukodystrophy and subacute sclerosing panencephalitis.

Additional neurologic diseases which can be treated or detected withmodified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention includecerebrovascular disorders (such as carotid artery diseases which includecarotid artery thrombosis, carotid stenosis and Moyamoya Disease),cerebral amyloid angiopathy, cerebral aneurysm, cerebral anoxia,cerebral arteriosclerosis, cerebral arteriovenous malformations,cerebral artery diseases, cerebral embolism and thrombosis such ascarotid artery thrombosis, sinus thrombosis and Wallenberg's Syndrome,cerebral hemorrhage such as epidermal hematoma, subdural hematoma andsubarachnoid hemorrhage, cerebral infarction, cerebral ischemia such astransient cerebral ischemia, Subclavian Steal Syndrome andvertebrobasilar insufficiency, vascular dementia such as multi-infarctdementia, periventricular leukomalacia, vascular headache such ascluster headache and migraine.

Additional neurologic diseases which can be treated or detected withmodified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention include dementiasuch as AIDS Dementia Complex, presenile dementia such as Alzheimer'sDisease and Creutzfeldt-Jakob Syndrome, senile dementia such asAlzheimer's Disease and progressive supranuclear palsy, vasculardementia such as multi-infarct dementia, encephalitis which includeencephalitis periaxialis, viral encephalitis such asepidemicencephalitis, Japanese Encephalitis, St. Louis Encephalitis,tick-borne encephalitis and West Nile Fever, acute disseminatedencephalomyelitis, meningoencephalitis such as uveomeningoencephaliticsyndrome, Postencephalitic Parkinson Disease and subacute sclerosingpanencephalitis, encephalomalacia such as periventricular leukomalacia,epilepsy such as generalized epilepsy, which includes infantile spasms,absence epilepsy, myoclonic epilepsy which includes MERRF Syndrome,tonic-clonic epilepsy, partial epilepsy such as complex partialepilepsy, frontal lobe epilepsy and temporal lobe epilepsy,post-traumatic epilepsy, status epilepticus such as Epilepsia PartialisContinua, and Hallervorden-Spatz Syndrome.

Additional neurologic diseases which can be treated or detected withmodified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention includehydrocephalus such as Dandy-Walker Syndrome and normal pressurehydrocephalus, hypothalamic diseases such as hypothalamic neoplasms,cerebral malaria, narcolepsy which includes cataplexy, bulbarpoliomyelitis, cerebripseudo tumor, Rett Syndrome, Reye's Syndrome,thalamic diseases, cerebral toxoplasmosis, intracranialtuberculoma andZellweger Syndrome, central nervous system infections such as AIDS,Dementia Complex, Brain Abscess, subdural empyema, encephalomyelitissuch as Equine Encephalomyelitis, Venezuelan Equine Encephalomyelitis,Necrotizing Hemorrhabaic Encephalomyelitis, Visna, and cerebral malaria.

Additional neurologic diseases which can be treated or detected withmodified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention include meningitissuch as arachnoiditis, aseptic meningitis such as viral meningitis whichincludes lymphocytic chronic meningitis, Bacterial meningitis whichincludes Haemophilus Meningitis, Listeria Meningitis, MeningococcalMeningitis such as Waterhouse-Fridericlisen Syndrome, PneumococcalMeningitis and meningeal tuberculosis, fungal meningitis such asCryptococcal Meningitis, subdural effusion, meningencephalitis, myelitissuch as transverse myelitis, neurosyphilis such as tabes dorsalis,poliomyelitis which includes bulbar poliomyelitis and post poliomyelitissyndrome, prion diseases (such as Creutzfeldt-Jakob Syndrome, BovineSpongiform Encephalopathy, Gerstmann-Straussler Syndrome, Kuru,Scrapie), and cerebral toxoplasmosis.

Additional neurologic diseases which can be treated or detected withmodified fusion proteins of the invention and/or polynucleotidesencoding transferrin fusion proteins of the invention include centralnervous system neoplasms such as brain neoplasms that includecerebellarneoplasms such as infratentorial neoplasms, cerebral ventricleneoplasms such as choroids plexus neoplasms, hypothalamic neoplasms andsupratentorial neoplasms, meningealneoplasms, spinal cord neoplasmswhich include epidural neoplasms, demyelinating diseases such as CanavanDiseases, diffuse cerebral sculleries which includesadrenoleukodystrophy, encephalitis periaxialis, globoid cellleukodystrophy, diffuse cerebral sclerosis such as metachromaticleukodystrophy, allergic encephalomyelitis, necrotizing hemorrhagicencephalomyelitis, progressive multifocal leukoencephalopathy, inmultiple sclerosis, central pontine myelinolysis, transverse myelitis,neuromyelitis optica, Scrapie, Swayback, Chronic Fatigue Syndrome,Visna, High Pressure Nervous Syndrome, Meningism, spinal cord diseasessuch as amyotonia congenita, amyotrophic lateral-sclerosis, spinalmuscular atrophy such as Werdnig-Hoffmann Disease, spinal cordcompression, spinal cord neoplasms such as epidural neoplasms,syringomyelia., Tabes Dorsalis, Stiff-Man Syndrome, mental retardationsuch as Angelman Syndrome, Cri-du-Chat Syndrome, De Lange's Syndrome,Down Syndrome, Gangliosidoses such as gangliosidoses G(MI), SandhoffDisease, Tay-Sachs Disease, Hartnup Disease, homocystinuria,Laurence-Moon-Bied Syndrome, Lesch-Nylian Syndrome, Maple Syrup UrineDisease, mucolipidosis such as fucosidosis, neuronalceroid-lipofuscinosis, oculocerebrorenal syndrome, phenylketonuria suchas maternal phenylketonuria, Prader-Willi Syndrome, Rett Syndrome,Rubinstein-Taybi Syndrome, Tuberous Sclerosis, WAGR Syndrome, nervoussystem abnormalities such as holoprosencephaly, neural tube defects suchas anencephaly which includes hydrangencephaly, Arnold-Chairi Deformity,encephalocele, meningocele, meningomyelocele, spinal dysraphism such asSpina bifida cystica and spina bifida occulta.

Endocrine system and/or hormone imbalance and/or diseases encompassdisorders of uterine motility including, but not limited tocomplications with pregnancy and labor (e.g., pre-term labor, post-termpregnancy, spontaneous abortion, and slow or stopped labor); anddisorders and/or diseases of the menstrual cycle, (e.g., dysmenorrheaand endometriosis).

Endocrine system and/or hormone imbalance disorders and/or diseasesinclude disorders and/or diseases of the pancreas, such as, for example,diabetes mellitus, diabetes insipidus, congenital pancreatic agenesis,pheochromocytoma islet cell tumor syndrome; disorders and/or diseases ofthe adrenal glands such as, for example, Addison's Disease,corticosteroid deficiency, virilizing disease, hirsutism, Cushing'sSyndrome, hyperaldosterlonism, pheochromocytoma; disorders and/ordiseases of the pituitary gland, such as, for example, hyperpituitarism,hypopituitarism, pituitary dwarfism, pituitary adenoma,panhypopituitarism, acromegaly, gigantism; disorders and/or diseases ofthe thyroid, including but not limited to, hyperthyroidism,hypothyroidism, Plummer's disease, Graves' disease (toxic diffusegoiter), toxic nodular goiter, thyroiditis (Hashimoto's thyroiditis,subacute granulomatous thyroiditis, and silent lymphocytic thyroiditis),Pendred's syndrome, myxedema, cretinism, thyrotoxicosis, thyroid hormonecoupling defect, thymic aplasia, Hurthle cell tumors of the thyroid,thyroid cancer, thyroid carcinoma, Medullary thyroid carcinoma;disorders and/or diseases of the parathyroid, such as, for example,hyperparathyroidism, hypoparathyroidism; disorders and/or diseases ofthe hypothalamus.

In addition, endocrine system and/or hormone imbalance disorders and/ordiseases may also include disorders and/or diseases of the testes orovaries, including cancer. Other disorders and/or diseases of the testesor ovaries further include, for example, ovarian cancer, polycysticovary syndrome, Klinefelter's syndrome, vanishing testes syndrome(bilateral anorchia), congenital absence of Leydig's cells,cryptorchidism, Noonan's syndrome, myotonic dystrophy, capillaryhaemangioma of the testis (benign), neoplasias of the testis andneotestis.

Moreover, endocrine system and/or hormone imbalance disorders and/ordiseases may also include disorders and/or diseases such as, forexample, polyglandular deficiency syndromes, pheochromocytoma,neuroblastoma, multiple Endocrine neoplasia, and disorders and/orcancers of endocrine tissues.

The modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay be used for the diagnosis, treatment, or prevention of diseasesand/or disorders of the reproductive system. Reproductive systemdisorders that can be treated by the compositions of the invention,include, but are not limited to, reproductive system injuries,infections, neoplastic disorders, congenital defects, and diseases ordisorders will result in infertility, complications with pregnancy,labor, or parturition, and postpartum difficulties.

Reproductive system disorders and/or diseases include diseases and/ordisorders, of the testes, including testicular atrophy, testicularfeminization, cryptorchism (unilateral and bilateral), anorchia, ectopictestis, epididymitis and orchitis (typically resulting from infectionssuch as, for example, gonorrhea, mumps, tuberculosis, and syphilis),testiculartorsiori, vasitis nodosa, germ cell tumors (e.g., seminomas,embryonal cell carcinomas, teratocarcinomas, choriocarcinomas, yolk sactumors, and teratomas), stromal tumors (e.g., Leydig cell tumors),hydrocele, hematocele, varicocele, spermatocele, inguinal hernia, anddisorders of sperm production (e.g. immotile cilia syndrome, spermia,asthenozoospermia, azoospermia, oligospermia, and teratozoospermia).

Reproductive system disorders also include disorders of the prostategland, such as acute non-bacterial prostatitis, chronic non-bacterialprostatitis, acute bacterial prostatitis, chronic bacterial prostatitis,postatodystonia, prostatosis, granulomatotis prostatitis, malacoplakia,benign prostatic hypertrophy or hyperplasia, and prostate neoplasticdisorders, including adenocarcinomas, transitional cell carcinomas,ductal carcinomas, and squamous cell carcinomas.

Additionally, the compositions of the invention may be useful in thediagnosis, treatment, and/or prevention of disorders or diseases of thepenis and urethra, including inflammatory disorders, such asbalanopbsthitis, balanitis xerotica obliterans, phimosis, paraphimosis,syphilis, herpes simplex virus, gonorrhea, non-gonococcal urethritis,chlamydia, mycoplasma, trichomonas, HIV, AIDS; Reiter's syndrome,condyloma acuminatum, condyloma latum, and pearly penile papules,urethral abnormalities, such as hypospadias, epispadias, and phimosis,premalignant lesions, including Erythroplasia of Queyrat, Bowen'sdisease, Bowenoid paplosis, criant condyloma of Buscke-Lowenstein, andvarrucous carcinoma; penile cancers, including squamous cell carcinomas,carcinoma in situ, verrucous carcinoma, and disseminated penilecarcinoma; urethral neoplastic disorders, including penile urethialcarcinoma, bulbomembranotis urethial carcinoma, and prostaticurethralcarcinoma; and erectile disorders, such as priapism, Peyronie's disease,erectile dysfunction, and impotence.

Moreover, diseases and/or disorders of the vas deferens includevasculititis and CBAVD (congenital bilateral absence of the vasdeferens); additionally, the transferrin fusion proteins of theinvention and/or polynucleotides encoding transferrin fusion proteins ofthe invention may be used in the diagnosis, treatment, and/or preventionof diseases and/or disorders of the seminal vesicles, including hydatiddisease, congenital chloride diarrhea, and polycystic kidney disease.

Other disorders and/or diseases of the male reproductive system include,for example, Klinefelters syndrome, Young's syndrome, prematureejaculation, diabetes mellitus, cystic fibrosis, Kartagener's syndrome,high fever, multiple sclerosis, and gynecomastia.

Further, the polynucleotides, modified fusion proteins of the inventionand/or polynucleotides encoding transferrin fusion proteins of theinvention may be used in the diagnosis treatment and/or prevention ofdiseases and/or disorders of the vagina and vulva, including bacterialvaginosis, candida vaginitis, herpes simplex virus, chancroid, granulomainguinale, lymphogranuloma venereum, scabies, human papillomavirus,vaginal trauma, vulvartrauma, adenosis, chlamydia vaginitis, gonorrhea,trichomonas vaginitis, condylomaacuminatum, syphilis, molluscumcontagiosum, atrophic vaginitis, Paaet's disease, lichensclerosus,lichen planus, vulvodynia, toxic shock syndrome, vaginismus,vulvovaginitis, vulvar vestibulitis, and neoplastic disorders, such assquamous cell hyperplasia, clear cell carcinoma, basal cell carcinoma,melanomas, cancer of Bartholin's gland, and vulvarintraepaelialneoplasia.

Disorders and/or diseases of the uterus include dysmenorrhea,retroverted uterus, endometriosis, fibroids, adenomyosis, anovulatorybleeding, amenorrhea, Cushiner's syndrome, hydatidiform moles,Asherman's syndrome, premature menopause, precocious puberty, uterinepolyps, dysfunctional uterine bleeding (e.g., due to aberrant hormonalsignals), and neoplastic disorders, such as adenocarcinomas,keiomyosarcomas, and sarcomas. Additionally, the transferrin fusionproteins of the invention and/or polynucleotides encoding transferrinfusion proteins of the invention may be useful as a marker or detectorof, as well as, in the diagnosis, treatment, and/or prevention ofcongenital uterine abnormalities, such as bicornuate uterus, septateuterus, simple unicomuate uterus, unicomuate uterus with a noncavitaryrudimentary horn, unicorriuate uterus with a non-communicating cavitaryrudimentary horn, unicomuate uterus with a communicating cavitary horn,arcuate uterus, uterine didelfus, and T-shaped uterus.

Ovarian diseases and/or disorders include an ovulation, polycystic ovarysyndrome (Stein-Leventhal syndrome), ovarian cysts, ovarianhypofunction, ovarian insensitivity to gonadotropins, ovarian overproduction of androgens, right ovarian vein syndrome, in amenorrhea,hirutism, and ovarian cancer (including, but not limited to, primary andsecondary cancerous growth, Sertoli-Leydig tumors, endometriod carcinomaof the ovary, ovarian papillary serous adenocarcinoma, ovarian mucinousadenocarcinoma, and Ovarian Krukenberg tumors).

Cervical diseases and/or disorders include cervicitis, chroniccervicitis, mucopurulent cervicitis, and cervical dysplasia, cervicalpolyps, Nabothian cysts, cervical erosion, cervical incompetence, andcervical neoplasms (including, for example, cervical carcinoma, squamousmetaplasia, squamous cell carcinoma, adenosquamous cell neoplasia, andcolumnar cell neoplasia).

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventioncan be used to treat or detect infectious agents. For example, byincreasing the immune response, particularly increasing theproliferation and differentiation of B and/or T cells, infectiousdiseases may be treated. The immune response may be increased by eitherenhancing an existing immune response, or by fusion proteins of theinvention and/or initiating a new immune response. Alternatively,polynucleotides encoding transferrin fusion proteins of the inventionmay also directly inhibit infectious agent, without necessarilyeliciting an immune response.

Viruses are one example of an infectious agent that can cause disease orsymptoms that can be treated or detected by transferrin fusion proteinsof the invention and/or polynucleotides encoding transferrin fusionproteins of the invention. Examples of viruses, include, but are notlimited to the following DNA and RNA viruses and viral families:Arbovirus, Adenoviridae, Arenaviridae, Arterivirus, Bimaviridae,Bunyaviridae, Caliciviridae, Circoviridae, Coronaviridae, Dengue, EBV,HIV, Flaviviridae, Hepadnaviridae Hepatitis, Herpesviridae (such as,Cytomegalovirus, Herpes Simplex, Herpes Zoster), Mononegavirus (e.g.,Paramyxoviridae, Morbillivirus, Rhabdoviridae), Orthomyxoviridae (e.g.,Influenza A, Influenza B, and parainfluenza), Papilloma virus,Papovaviridae, Parvoviridaes, Picornaviridae, Poxviridae (such asSmallpox or Vaccinia), Reoviridae (e.g., Rotavirus), Retroviridae(HTLV-I, HTLV-11, -Lentivirus), and Togaviridae (e.g., Rubivirus).

Similarly, bacterial and fungal agents that can cause disease orsymptoms that can be treated or detected by transferrin fusion proteinsof the invention and/or polynucleotides encoding transferrin fusionproteins of the invention include, but not limited to, the followingGram-negative and Gram-positive bacteria, bacterial families, and fungi:Actinomyces (e.g., Norcardia), Acinetobacter, Cryptococcus neoformans,Aspergillus, Bacillaceae (e.g., Bacillus anthrasis), Bacteroides (e.g.,Bacteroides fragilis), Blastomycosis, Bordetella, Borrelia (e.g.,Borrelia burgdorferi) Brucella, Candidia, Campylobacter, Chlamydia,Clostridiuffi (e.g., Clostridium botulinum, Clostridium difficile,Clostridium perfringens, Clostridiumtetani), Coccidioides,Corynebacterium (e.g., Corynebacterium-diptheriae), Cryptococcus,Dermatocycoses, E. coli (e.g., Enterotoxigenic E. coli andEnterohemorrhagic E. coli), Enterobacter (e.g. Enterobacter aerogenes),Enterobacteriaceae (Klebsiella, Salmonella (e.g., Salmonella typhi,Salmonella enteritidis, Salmonella typhi), Serratia, Yersinia,Shigella), Erysipelothrix, Haemophilus (e.g., Haemophilus influenza typeB), Helicobacter, Legionella (e.g., Legionella pneumophila), Leptospira,Listeria (e.g., Listeria monocytogenes), Mycoplasma, Mycobacterium(e.g., Mycobacterium leprae and Mycobacterium tuberculosis), Vibrio(e.g., Vibrio cholerae), Neisseriaceae (Neisseriagonorrhea, Neisseriameningitidis), Pasteurellaceae, Proteus, Pseudomonas (e.g., Pseudomionasaeruginosa), Rickettsiaceae, Spirochetes (e.g., Treponenza. spp.,Leptospiraspp., Borrielia spp), Shigella spp., Staphylococcus (e.g.,Staphylococcus aureus), Meningiococcus, Pneumococcus and Streptococcus(e.g., Streptococcus pneumoniae and Groups A, B, and C Streptococci),and Ureaplasmas.

Moreover, parasitic agents causing disease or that can be treated,prevented, and/or diagnosed by fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventioninclude, but not limited to, the following families or class: Amebiasis,Babesiosis, Coccidiosis, Cryptosporidiosis, Dientamoebiasis, Dourine,Ectoparasitic, Giardias, Helminthiasis, Leishmaniasis, Schistisoma,Theileriasis, Toxoplasmosis, Trypanosomiasis, and Trichomonas andSporozoans (e.g., Plasmodium vivax, Plasmodium falciparium, Plasmodiummalariae and Plasmodium ovale).

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventioncan be used to differentiate, proliferate, and attract cells, pleadingto the regeneration of tissues. (See, Science 276:59-87 (1997)). Theregeneration of tissues could be used to repair, replace, or protecttissue damaged by congenital defects, trauma (wounds, burns, incisions,or ulcers), age, disease (e.g. osteoporosis, osteocarthritis,periodontal disease, liver failure), surgery, including cosmetic plasticsurgery, fibrosis, reperfusion injury, or systemic cytokine damage.

Tissues that could be regenerated using the present invention includeorgans (e.g., pancreas, liver, intestine, kidney, skin, endothelium),muscle (smooth, skeletal or cardiac), vasculature (including vascularand lymphatics), nervous, hematopoietic, and skeletal (bone, cartilage,tendon, and ligament) tissue. Preferably, regeneration occurs without ordecreased scarring. Regeneration also may include angiogenesis.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the invention,may be used to treat, prevent, diagnose, and/or prognosegastrointestinal disorders, including inflammatory diseases and/orconditions, infections, cancers (e.g., intestinal neoplasms (carcinoidtumor of the small intestine, non-Hodgkin's lymphoma of the smallintestine, small bowel lymphoma), and ulcers, such as peptic ulcers.

Gastrointestinal disorders include dysphagia, odynophagia, inflammationof the esophagus, peptic esophagitis, gastric reflux, submucosalfibrosis and structuring, Mallory-Weiss lesions, lipomas, epidennalcancers, adeoncarcinomas, gastric retention disorders, gastroenteritis,gastric atrophy, gastric/stomach cancers, polyps of the stomach,autoimmune disorders such as pernicious anemia, pyloric stenosis,gastritis (bacterial, viral, eosinophilic, stress-induced, chronicerosive, atrophic, plasma cell, and Menetrier's), and peritonealdiseases (e.g., chylo perioneum, hemoperitoneum, mesenteric cyst,mesentericlymphadenitis, mesenteric vascular occlusion, panniculiti,neoplasms, peritonitis, pneumoperitoneum, bubphrenic abscess.

Gastrointestinal disorders also include disorders associated with thesmall intestine, such as malabsorption syndrome's, distension, irritablebowel syndrome, sugar intolerance, celiac disease, duodenal ulcers,duodenitis, tropical sprue, Whipple's disease, intestinallymphangiectasia, Crohn's disease, appendicitis, obstructions of theileum, Meckel's diverticulum, multiple diverticula, failure of completerotation of the small and large intestine, lymphoma, and bacterial andparasitic diseases (such as Traveler's diarrhea, typhoid andparatyphoid, cholera, infection by Roundworms (Ascariasis lumbricoides),Hookworms (Ancylostoma duodenale), Threadworms (Enterobiusvermicularis), Tapeworms Taenia saginata, Echinococcus granulosus,Diphyllobothrium spp. and T. solium).

Liver diseases and/or disorders include intrahepatic cholestasis(Alagille syndrome, biliary liver cirrhosis), fatty, liver (alcoholicfatty liver, Reye's syndrome), hepatic veiri, thrombosis,hepatolentricular degeneration, hepatomegaly, hepatopulmonary syndrome,hepatorenal, syndrome, portal hypertension (esophageal and gastricvarices), liver abscess (amebic liver abscess), liver cirrhosis(alcoholic, biliary and experimental), alcoholic liver diseases (fattyliver, hepatitis, cirrhosis), parasitic (hepatic echinococcosis,fascioliasis, amebic liver abscess), jaundice (hemolytic,hepatocellular, and cholestatic), cholestasis, portal hypertension,liver, enlargement, ascites, hepatitis (alcoholic hepatitis,intra-familial hepatitis, chronic hepatitis (autoimmune, hepatitis B,hepatitis C, hepatitis D, drug induced), toxic hepatitis, viral humanhepatitis (hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitisE), Wilson's disease, granulomatous hepatitis, secondary biliarycirrhosis, hepaticencephalopathy, portal hypertension, varices, hepaticencepbalopathy, primary biliary hemangiomas, bilecirrhosis, primarysclerosing cholangitis, hepatocellular adenoma, stones, liver failure(hepatic encephalopathy, acute liver failure), and liver neoplasms(ancriomyolipoma, calcified liver metastases, cystic liver metastases,epithelial tumors, fibro lamellar hepatocarcinoma, focal nodularhyperplasia, hepatic adenoma, hepatobiliarycystadenoma, hepatoblastoma,hepatocellular carcinoma, hepatoma, liver cancer, liverhemangioendothelioma, mesenchymal hamartoma, mesenchymal tumors ofliver, nodular regenerative hyperplasia, benign liver tumors (Hepaticcysts, Simple cysts, Polycystic liver disease, Hepatobiliarycystadenoma, Choledochal cysts, Mesenchymal tumors, Mesenchymalhamartoma, Infantile hemangioendothelioma, Hemangioma, Peliosis hepatis,Lipomas, Inflammatory pseudo tumor, Miscellaneous Epithelial tumors,Bile duct epithelium (Bile duct hamartoma, Bile duct adenoma),Hepatocyte (Adenoma, Focal nodular hyperplasia, Nodular regenerativehyperplasia), malignant liver tumors (hepatocellular, hepatoblastoma,hepatocellular carcinoma, cholangiocellular, cholangiocarcinoma,cystadenocarcinoma, tumors of blood vessels, anaiosarcoma, Karposi'ssarcoma, hemangioendothelioma, other tumors, embryonal sarcorria,fibrosarcoma, rhabdomyosarcoma, carcinosarcoma, teratoma, carcinoid,squamous carcinoma, primarylymphorria)), peliosis hepatis,erythrohepatic porphyria, hepatic porphyria (acuteinterirtittentporphyria; porphyria cutanea tarda), Zelli Negersyndrome).

Pancreatic diseases and/or disorders include acute pancieatitis, chronicpancreatitis (acute necrotizing pancreatitis, alcoholic pancreatitis),neoplasins (adenocarcinoma of the pancreas, cystadenocarcinoma,insulinoma, gastrinoma, and glucacronoma, cysticcitmeoplasms, islet-celltumors, pancreoblastoma), and other pancreatic diseases (e.g.,cysticfibrosis, cyst (pancreatic pseudocyst, pancreatic fistula,insufficiency)).

Gallbladder diseases include gallstones (cholelithiasis andcholedocholithiasis), postcholeeystectomy syndrome, diverticulosis ofthe gallbladder, acute cholecystitis, chronic cholecystitis, bile ducttumors, and mucocele.

Diseases and/or disorders of the large intestine includeantibiotic-associated colitis, diverticulitis, ulcerative colitis,acquired megacolon, abscesses, fungal and bacterial infections,anorectal disorders (e.g., fissures, hemorrhoids), colonic diseases(colitis, colonic neoplastris, colon cancer, adenomatous colon polyps(e.g., villous adenoma), coloncarcinoma, colorectal cancer, colonicdiverticulitis, colonic diverticulosis, megacolon, Hirschsprung disease,toxic megacolon, sigmoid diseases proctocolitis, sigmoinneoplasmsj,constipation, Crohn's disease, diarrhea (infantile diarrhea, dysentery),duodenal diseases (duodenal neoplasins, duodenal obstruction, duodenalulcer, duodenitis), enteritis (enterocolitis), HIV enteropathy, lealdiseases (leal neoplasins, ileitis), immunoproliferative smallintestinal disease, inflammatory bowel disease (ulcerative colitis,Crohn's disease), intestinal atresia, parasitic diseases (anisakiasis,balantidiasis, blastocystis infections, cryptosporidiosis,dientamoebiasis, amebic dysentery, giardiasis), intestinal fistula(rectal fistula), intestinal neoplasms (cecal neoplasms, colonicneolasms, duodenalpneoplasms, leal neoplasms, intestinal polyps, jejunalneoplasins, rectal neoplasms), intestinal obstruction (afferent loopsyndrome, duodenal obstruction, impacted feces, intestinal pseudoobstruction cecal volvulus, intussusception), intestinal perforation,intestinal polyps (colonic polyps, gardner syndrome, peutz-jegherssyndrome), jejunal diseases Oejunal neoplasms), mal absorption syndromes(blind loop syndrome, celiac disease, lactose intolerance, short bowlsyndrome, tropical sprue, whipple's disease), mesenteric vascularocclusion, pneumatosis cystoides intestinalis, protein losingenteropathies (intestinal lymphagiectasis), rectal diseases (anusdiseases, fecal incontinence, hemorrhoids, proctitis, rectal fistula,rectal prolapse, rectocele), peptic ulcer (duodenalulcer, pepticesophagitis, hemorrhage, perforation; stomach ulcer, Zollinger-Ellisonsyndrome), postgastrectomy syndromes (dumping syndrome), stomachdiseases (e.g., achlorhydria, duodenogastric reflux (bile reflux),gastric antral vascular ectasia, gastricfistula, gastric outletobstruction, gastritis (atrophic or hypertrophic), gastroparesis,stomach dilatation; stomach diverticulum, stomach neoplasms (gastriccancer, gastric polyps, gastric adenocarcinoma, hyperplastic gastricpolyp), stomach rupture, stomach ulcer, stomach volvulus), tuberculosis,visceroptosis, vomiting (e.g., hematemesis, hyperemesisgravidanim,postoperative nausea and vomiting) and hemorrhagic colitis.

Further diseases and/or disorders of the gastrointestinal system includebiliary tract diseases, such as, gastroschisis, fistula (e.g., biliaryfistula, esophageal fistula, gastricfistula, intestinal fistula,pancreatic fistula), neoplasms (e.g., biliary tract neoplasins,esophageal neoplasms; such as adenocarcinoma of the esophagus,esophageal squamous cell carcinoma, gastrointestinal neoplasms,pancreatic neoplasins, such as adenocarcinoma of the pancreas, mucinouscystic neoplasm of the pancreas, pancreatic eystic neoplasms,pancreatoblastoma, and peritoneal neoplasms), esophageal disease (e.g.,bullous diseases, candidiasis, glycoaenie acanthosis, ulceration,barrett esophagus varices, atresia, cyst, diverticulum. (e.g., Zenker'sdiverticulum), fistula (e.g., tracheoesophageal fistula), motilitydisorders (e.g., CREST syndrome, deglutition disorders, achalasia,spasm, gastroesophageal reflux), neoplasms, perforation (e.g., Boerhaavesyndrome, Mallory-Weiss syndrome), stenosis, esophagitis, diaphragmatichernia (e.g., hiatal hernia); gastrointestinal diseases, such as,gastroenteritis (e.g., cholera morbus, norwalk virus infection),hemorrhage (e.g., hematemesis, melena, peptic ulcer hemorrhage), stomachneoplasms (gastric cancer, gastric polyps, gastric adenocarcinoma,stomach cancer)), hernia (e.g., congenital diaphragmatic hernia, femoralhernia, inguinal hernia, obturator hernia, umbilical hernia, ventralhernia), and intestinal diseases (e.g., cecal diseases (appendicitis,cecal neoplasms)).

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay have chemotaxis activity. A chemotaxic molecule attracts ormobilizes cells (e.g., monocytes, fibroblasts, neutrophils, T-cells,mast cells, eosinophils, epithelial and/or endothelial cells) to aparticular site in the body, such as inflammation, infection, or site ofhyperproliferation. The mobilized cells can then fight off and/or healthe particular trauma or abnormality.

Modified transferrin fusion proteins of the invention and/orpolynucleotides encoding transferrin fusion proteins of the inventionmay increase chemotaxic activity of particular cells. These chemotacticmolecules can then be used to treat inflammation, infection,hyperproliferative disorders, or any immune system disorder byincreasing the number of cells targeted to a particular location in thebody.

Oral Pharmaceutical Compositions and Delivery Methods

In the present invention, Tf fusion proteins, including but not limitedto modified Tf fusion proteins, may be formulated for oral, delivery. Inparticular, certain fusion proteins of the invention that are used totreat certain classes of diseases or medical conditions may beparticularly amenable for oral formulation and delivery. Such classes ofdiseases or conditions include, but are not limited to, acute, chronicand recurrent diseases. Chronic or recurrent diseases include, but arenot limited to, viral disease or infections, cancer, a metabolicdiseases, obesity, autoimmune diseases, inflammatory diseases, allergy,graft-vs.-host disease, systemic microbial infection, anemia,cardiovascular disease, psychosis, genetic diseases, neurodegenerativediseases, disorders of hematopoietic cells, diseases of the endocrinesystem or reproductive systems, gastrointestinal diseases. Examples ofthese classes of disease include diabetes, multiple sclerosis, asthma,HCV or HIV infections, hypertension, hypercholesterolemia, arterialscherosis, arthritis, and Alzheimer's disease. In many chronic diseases,oral formulations of Tf fusion proteins of the invention and methods ofadministration are particularly useful because they allow long-termpatient care and therapy via home oral administration without relianceon injectable treatment or drug protocols.

Oral formulations and delivery methods comprising Tf fusion proteins ofthe invention take advantage of, in part, transferrin receptor mediatedtranscytosis across the gastrointestinal (GI) epithelium. The Tfreceptor is found at a very high density in the human GI epithelium,transferrin is highly resistant to tryptic and chymotryptic digestionand Tf chemical conjugates have been used to successfully deliverproteins and peptides across the GI epithelium (Xia et al., (2000) J.Pharmacol. Experiment. Therap., 295:594-600; Xia et al. (2001)Pharmaceutical Res., 18(2):191-195; and Shah et al. (1996) J.Pharmaceutical Sci., S5(12):1306-1311, all of which are hereinincorporated by reference in their entirety). Once transported acrossthe GI epithelium, Tf fusion proteins of the invention exhibit extendedhalf-life in serum, that is, the therapeutic protein or peptide(s)attached or inserted into Tf exhibit an extended serum half-lifecompared to the protein or peptide in its non-fused state.

Oral formulations of Tf fusion proteins of the invention may be preparedso that they are suitable for transport to the GI epithelium andprotection of the Tf fusion protein component and other activecomponents in the stomach. Such formulations may include carrier anddispersant components and may be in any suitable form, includingaerosols (for oral or pulmonary delivery), syrups, elixirs, tablets,including chewable tablets, hard or soft capsules, troches, lozenges,aqueous or oily suspensions, emulsions, cachets or pellets granulates,and dispersible powders. Preferably, Tf fusion protein formulations areemployed in solid dosage forms suitable for simple, and preferably oral,administration of precise dosages. Solid dosage forms for oraladministration are preferably tablets, capsules, or the like.

For oral administration in the form of a tablet or capsule, care shouldbe taken to ensure that the composition enables sufficient activeingredient to be absorbed by the host to produce an effective response.Thus, for example, the amount of Tf fusion protein may be increased overthat theoretically required or other known measures such as coating orencapsulation may be taken to protect the polypeptides from enzymaticaction in the stomach.

Traditionally, peptide and protein drugs have been administered byinjection because of the poor bioavailability when administerednon-parenterally, and in particular orally. These drugs are prone tochemical and conformational instability and are often degraded by theacidic conditions in the stomach, as well as by enzymes in the stomachand gastrointestinal tract. In response to these delivery problems,certain technologies for oral delivery have been developed, such asencapsulation in nanoparticles composed of polymers with a hydrophobicbackbone and hydrophilic branches as drug carriers, encapsulation inmicroparticles, insertion into liposomes in emulsions, and conjugationto other molecules. All of which may be used with the Tf fusionmolecules of the present invention.

Examples of nanoparticles include mucoadhesive nanoparticles coated withchitosan and Carbopol (Takeuchi et al., Adv. Drug Deliv. Rev.47(1):39-54, 2001) and nanoparticles containing charged combinationpolyesters, poly(2-sulfobutyl-vinyl alcohol) andpoly(D,L-lactic-co-glycolic acid) (Jung et al., Eur. J. Pharm. Biopharm.50(1):147-160, 2000). Nanoparticles containing surface polymers withpoly-N-isopropylacrylamide regions and cationic poly-vinylamine groupsshowed improved absorption of salmon calcitonin when administered orallyto rats.

Drug delivery particles composed of alginate and pectin, strengthenedwith polylysine, are relatively acid and base resistant and can be usedas a carrier for drugs. These particles combine the advantages ofbioadhesion, enhanced absorption and sustained release (Liu et al., J.Pharm. Pharmacol. 51(2):141-149, 1999).

Additionally, lipoamino acid groups and liposaccharide groups conjugatedto the N- and C-termini of peptides such as synthetic somatostatin,creating an amphipathic surfactant, were shown to produce a compositionthat retained biological activity (Toth et al., J. Med. Chem.42(19):4010-4013, 1999).

Examples of other peptide delivery technologies include carbopol-coatedmucoadhesive emulsions containing the peptide of interest and eithernitroso-N-acetyl-D,L-penicillamine and carbolpol or taurocholate andcarbopol. These were shown to be effective when orally administered torats to reduce serum calcium concentrations (Ogiso et al., Biol. Pharm.Bull. 24(6):656-661, 2001). Phosphatidylethanol, derived fromphosphatidylcholine, was used to prepare liposomes containingphosphatidylethanol as a carrier of insulin. These liposomes, whenadministered orally to rats, were shown to be active (Kisel et J. Pharm.216(1-2):105-114, 2001).

Insulin has also been formulated in poly(vinyl alcohol)-gel spherescontaining insulin and a protease inhibitor, such as aprotinin orbacitracin. The glucose-lowering properties of these gel spheres havebeen demonstrated in rats, where insulin is released largely in thelower intestine (Kimura et al., Biol. Pharm. Bull. 19(6):897-900, 1996.

Oral delivery of insulin has also been studied using nanoparticles madeof poly(alkyl cyanoacrylate) that were dispersed with a surfactant in anoily phase (Damge et al., J. Pharm. Sci. 86(12):1403-1409, 1997) andusing calcium alginate beads coated with chitosan (Onal et al., Artif.Cells Blood Substit. Immobil. Biotechnol. 30(3):229-237, 2002).

In other methods, the N- and C-termini of a peptide are linked topolyethylene glycol and then to allyl chains to form conjugates withimproved resistance to enzymatic degradation and improved diffusionthrough the GI wall (www.nobexcorp.com).

BioPORTER® is a cationic lipid mixture, which interacts non-covalentlywith peptides to create a protective coating or layer. The peptide-lipidcomplex can fuse to the plasma membrane of cells, and the peptides areinternalized into the cells (www.genetherapysystems.com).

In a process using liposomes as a starting material, cochleate-shapedparticles have been developed as a pharmaceutical vehicle. A peptide isadded to a suspension of liposomes containing mainly negatively chargedlipids. The addition of calcium causes the collapse and fusion of theliposomes into large sheets composed of lipid bilayers, which thenspontaneously roll up or stack into cochleates (U.S. Pat. No. 5,840,707;http://www.biodeliverysciences.com).

Compositions comprising Tf fusion protein intended for oral use may beprepared according to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions may contain one ormore agents selected from the group consisting of sweetening agents inorder to provide a pharmaceutically elegant and palatable preparation.For example, to prepare orally deliverable tablets, Tf fusion protein ismixed with at least one pharmaceutical excipient, and the solidformulation is compressed to form a tablet according to known methods,for delivery to the gastrointestinal tract. The tablet composition istypically formulated with additives, e.g. a saccharide or cellulosecarrier, a binder such as starch paste or methyl cellulose, a filler, adisintegrator, or other additives typically usually used in themanufacture of medical preparations. To prepare orally deliverablecapsules, DHEA is mixed with at least one pharmaceutical excipient, andthe solid formulation is placed in a capsular container suitable fordelivery to the gastrointestinal tract. Compositions comprising Tffusion protein may be prepared as described generally in Remington'sPharmaceutical Sciences, 18th Ed. 1990 (Mack Publishing Co. Easton Pa.18042) at Chapter 89, which is herein incorporated by reference.

As described above, many of the oral formulations of the invention maycontain inert ingredients which allow for protection against the stomachenvironment, and release of the biologically active material in theintestine. Such formulations, or enteric coatings, are well known in theart. For example, tablets containing Tf fusion protein in admixture withnon-toxic pharmaceutically acceptable excipients which are suitable formanufacture of tablets may be used. These excipients may be inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, maize starch, gelatin or acacia, and lubricating agents,for example, magnesium stearate, stearic acid, or talc.

The tablets may be uncoated or they may be coated with known techniquesto delay disintegration and absorption in the gastrointestinal track andthereby provide a sustained action over a longer period of time. Forexample, a time delay material such as glyceryl monostearate or glyceryldistearate alone or with a wax may be employed.

Formulations for oral use may also be presented as hard gelatin capsulesWherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate, or kaolin or as softgelatin capsules wherein the active ingredient is mixed with an aqueousor an oil medium, for example, arachis oil, peanut oil, liquid paraffinor olive oil.

Aqueous suspensions may contain Tf fusion protein in the admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example, sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally occurring phosphatide,for example, lecithin, or condensation products of an alkylene oxidewith fatty acids, for example, polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample, heptadecylethyloxycetanol, or condensation products of ethyleneoxide with partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyoxyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives for example,ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one ormore flavoring agents and one or more sweetening agents such as sucroseor saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example, arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilsuspensions may contain a thickening agent, for example, beeswax, hardparaffin or cetyl alcohol. Sweetening agents, such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of anantioxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient andadmixture with dispersing or wetting agent, suspending agent and one ormore preservatives. Suitable dispersing or wetting agents and suspendingagents are exemplified by those already mentioned above. Additionalexcipients, for example, sweetening, flavoring and coloring agents, mayalso be present.

The pharmaceutical compositions containing Tf fusion protein may also bein the form of oil-in-water emulsions. The oil phase may be a vegetableoil, for example, olive oil or arachis oil, or a mineral oil forexample, gum acacia or gum tragacanth, naturally-occurring phosphotides,for example soybean lecithin, and esters or partial esters derived fromfatty acids and hexitol anhydrides, for example, sorbitan monooleate,and condensation products of the same partial esters with ethyleneoxide, for example, polyoxyethylene sorbitan monooleate. The emulsionsmay also contain sweetening and flavoring agents.

Syrups and elixirs containing Tf fusion protein may be formulated withsweetening agents, for example, glycerol, sorbitol or sucrose. Suchformulations may also contain a demulcent, a preservative and flavoringand coloring agents. The pharmaceutical compositions may be in the formof a sterile injectable preparation, for example, as a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparations may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvate, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this period any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

Pharmaceutical compositions may also be formulated for oral deliveryusing polyester microspheres, zein microspheres, proteinoidmicrospheres, polycyanoacrylate microspheres, and lipid-based systems(see, for example, DiBase and Morrel, Oral Delivery of MicroencapsulatedProteins, in Protein Delivery: Physical Systems, Sanders and Hendren(eds.), pages 255-288 (Plenum Press 1997)).

The proportion of pharmaceutically active Tf fusion protein to carrierand/or other substances may vary from about 0.5 to about 100 wt. %(weight percent). For oral use, the pharmaceutical formulation willgenerally contain from about 5 to about 100% by weight of the activematerial. For other uses, the formulation will generally have from about0.5 to about 50 wt. % of the active material.

Tf fusion protein formulations employed in the invention provide aneffective amount of Tf fusion protein upon administration to anindividual. As used in this context, an “effective amount” of Tf fusionis an amount that is effective to ameliorate a symptom of a disease.

The Tf fusion protein composition of the present invention may be,though not necessarily, administered daily, in an effective amount toameliorate a symptom. Generally, the total daily dosage will be at leastabout 50 mg, preferably at least about 100 mg, and more preferably atleast about 200 mg, and preferably not more than 500 mg per day,administered orally, e.g., in 4 capsules or tablets, each containing 50mg Tf fusion protein. Capsules or tablets for oral delivery canconveniently contain up to a full daily oral dose, e.g., 200 mg or more.

In a particularly preferred embodiment, oral pharmaceutical compositionscomprising Tf fusion protein are formulated in buffered liquid formwhich is then encapsulated into soft or hard-coated gelatin capsuleswhich are then coated with an appropriate enteric coating. For the oralpharmaceutical compositions of the invention, the location of releasemay be anywhere in the GI system, including the small intestine (theduodenum, the jejunum, or the ileum), or the large intestine.

In other embodiments, oral compositions of the invention are formulatedto slowly release the active ingredients, including the Tf fusionproteins of the invention, in the GI system using known delayed releaseformulations.

Tf fusion proteins of the invention for oral delivery are capable ofbinding the Tf receptor found in the GI epithelium. To facilitate thisbinding and receptor mediated transport, Tf fusion proteins of theinvention are typically produced with iron and in some instancescarbonate, bound to the Tf moiety. Processes and methods to load the Tfmoiety of the fusion protein compositions of the invention with iron andcarbonate are known in the art

In some pharmaceutical formulations of the invention, the Tf moiety ofthe Tf fusion protein may be modified to increase the affinity oravidity of the Tf moiety to iron. Such methods are known in the art. Forinstance, mutagenesis can be used to produce mutant transferrin moietiesthat bind iron more avidly than natural transferrin. In human serumtransferrin, the amino acids which are ligands for metal ion chelationinclude, but are not limited to N lobe amino acids Asp63, Tyr 95,Tyr188, Lys206, His207 and His249; and C lobe amino acids Asp392,Tyr426, Tyr517 and His5S5 of SEQ ID NO: 3 (the number beside the aminoacid indicates the position of the amino acid residue in the primaryamino acid sequence where the valine of the mature protein is designatedposition 1). See U.S. Pat. No. 5,986,067, which is herein incorporatedbe reference. In one embodiment, the Lys206 and His207 residues withinthe N lobe are replaced with Gln and Glu, respectively.

In some pharmaceutical formulations of the invention, the Tf fusionprotein is engineered to contain a cleavage site between the therapeuticprotein or peptide and the Tf moiety. Such cleavable sites or linkersare known in the art.

Pharmaceutical compositions of the invention and methods of theinvention may include the addition of a transcytosis enhancer tofacilitate transfer of the fusion protein across the GI epithelium. Suchenhancers are known in the art. See Xia et al., (2000) J. Pharmacol.Experiment. Therap., 295:594-600; and Xia et al. (2001) PharmaceuticalRes., 18(2):191-195.

In preferred embodiments of the invention, oral pharmaceuticalformulations include Tf fusion proteins comprising a modified Tf moietyexhibiting reduced or no glycosylation fused at the N terminal end to aninsulin or GLP-1 protein or peptide as described above. Suchpharmaceutical compositions may be used to treat glucose imbalancedisorders such as diabetes by oral administration of the pharmaceuticalcomposition comprising an effective dose of fusion protein.

The effective dose of fusion protein may be measured in a numbers ofways, including dosages calculated to alleviate symptoms associated witha specific disease state in a patient, such as the symptoms of diabetes.In other formulations, dosages are calculated to comprise an effectiveamount of fusion protein to induce a detectable change in blood glucoselevels in the patient. Such detectable changes in blood glucose mayinclude a decrease in blood glucose levels of between about 1% and 90%,or between about 5% and about 80%. These decreases in blood glucoselevels will be dependent on the disease condition being treated andpharmaceutical compositions or methods of administration may be modifiedto achieve the desired result for each patient. In other instances, thepharmaceutical compositions are formulated and methods of administrationmodified to detect an increase in the activity level of the therapeuticprotein or peptide in the patient, for instance, detectable increases inthe activities of insulin or GLP-1. Such formulations and methods maydeliver between about 1 pg to about 100 mg/kg body weight of fusionprotein, about 100 ng to about 100 μg/kg body weight of fusion protein,about 100 μg/to about 100 mg/kg body weight of fusion protein, about 1μg to about 1 g of fusion protein, about 10 μg to about 100 mg of fusionprotein or about 10 mg to about 50 mg of fusion protein. Formulationsmay also be calculated using a unit measurement of therapeutic proteinactivity, such as about 5 to about 500 units of human insulin or about10 to about 100 units of human insulin. The measurements by weight oractivity can be calculated using known standards for each therapeuticprotein or peptide fused to Tf.

The invention also includes methods of orally administering thepharmaceutical compositions of the invention. Such methods may include,but are not limited to, steps of orally administering the compositionsby the patient or a caregiver. Such administration steps may includeadministration on intervals such as once or twice per day depending onthe Tf fusion protein, disease or patient condition or individualpatient. Such methods also include the administration of various dosagesof the individual Tf fusion protein. For instance, the initial dosage ofa pharmaceutical composition may be at a higher level to induce adesired effect, such as reduction in blood glucose levels. Subsequentdosages may then be decreased once a desired effect is achieved. Thesechanges or modifications to administration protocols may be done by theattending physician or health care worker. In some instances, thechanges in the administration protocol may be done by the individualpatient, such as when a patient is monitoring blood glucose levels andadministering a mTf-insulin or mTf-GLP-1 oral composition of theinvention.

The invention also includes methods of producing oral compositions ormedicant compositions of the invention comprising formulating a Tffusion protein of the invention into an orally administerable form. Inother instances, the invention includes methods of producingcompositions or medicant compositions of the invention comprisingformulating a Tf fusion protein of the invention into a form suitablefor oral administration.

Moreover, the present invention includes pulmonary delivery of the Tffusion protein formulations. Pulmonary delivery is particularlypromising for the delivery of macromolecules which are difficult todeliver by other routes of administration. Such pulmonary delivery canbe effective both for systemic delivery and for localized delivery totreat diseases of the lungs, since drugs delivered to the lung arereadily absorbed through the alveolar region directly into the bloodcirculation.

The present invention provides compositions suitable for forming a drugdispersion for oral inhalation (pulmonary delivery) to treat variousconditions or diseases. The Tf fusion protein formulation could bedelivered by different approaches such as liquid nebulizers,aerosol-based metered dose inhalers (MDI's), and dry powder dispersiondevices. In formulating compositions for pulmonary delivery,pharmaceutically acceptable carriers including surface active agents orsurfactants and bulk carriers are commonly added to provide stability,dispersibility, consistency, and/or bulking characteristics to enhanceuniform pulmonary delivery of the composition to the subject.

Surface active agents or surfactants promotes absorption of polypeptidethrough mucosal membrane or lining. Useful surface active agents orsurfactants include fatty acids and salts thereof, bile salts,phospholipid, or an alkyl saccharide. Examples of fatty acids and saltsthereof include sodium, potassium and lysine salts of caprylate (C₈),caprate (C₁₀), laurate (C₁₂) and myristate (C₁₄). Examples of bile saltsinclude cholic acid, chenodeoxycholic acid, glycocholic acid,taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholicacid, deoxycholic acid, glycodeoxycholic acid, taurodeoxycholic acid,lithocholic acid, and ursodeoxycholic acid.

Examples of phospholipids include single-chain phospholipids, such aslysophosphatidylcholine, lysophosphatidylglycerol,lysophosphatidylethanolamine, lysophosphatidylinositol andlysophosphatidylserine; or double-chain phospholipids, such asdiacylphosphatidylcholines, diacylphosphatidylglycerols,diacylphosphatidylethanolamines, diacylphosphatidylinositols anddiacylphosphatidylserines. Examples of alkyl saccharides include alkylglucosides or alkyl maltosides, such as decyl glucoside and dodecylmaltoside.

Pharmaceutical excipients that are useful as carriers includestabilizers such as human serum albumin (HSA); bulking agents such ascarbohydrates, amino acids and polypeptides; pH adjusters or buffers;salts such as sodium chloride; and the like. These carriers may be in acrystalline or amorphous form or may be a mixture of the two.

Examples of carbohydrates for use as bulking agents includemonosaccharides such as galactose, D-mannose, sorbose, and the like;disaccharides, such as lactose, trehalose, and the like; cyclodextrins,such as 2-hydroxypropyl-.beta.-cyclodextrin; and polysaccharides, suchas raffinose, maltodextrins, dextrans, and the like; alditols, such asmannitol, xylitol, and the like. Examples of polypeptides for use asbulking agents include aspartame. Amino acids include alanine andglycine, with glycine being preferred.

Additives, which are minor components of the composition, may beincluded for conformational stability during spray drying and forimproving dispersibility of the powder. These additives includehydrophobic amino acids such as tryptophan, tyrosine, leucine,phenylalanine, and the like.

Suitable pH adjusters or buffers include organic salts prepared fromorganic acids and bases, such as sodium citrate, sodium ascorbate, andthe like; sodium citrate is preferred.

The Tf fusion compositions for pulmonary delivery may be packaged asunit doses where a therapeutically effective amount of the compositionis present in a unit dose receptacle, such as a blister pack, gelatincapsule, or the like. The manufacture of blister packs or gelatincapsules is typically carried out by methods that are generally wellknown in the packaging art.

U.S. Pat. No. 6,524,557 discloses a pharmaceutical aerosol formulationcomprising (a) a HFA propellant; (b) a pharmaceutically activepolypeptide dispersible in the propellant; and (c) a surfactant which isa C₈-C₁₆ fatty acid or salt thereof, a bile salt, a phospholipid, or analkyl saccharide, which surfactant enhances the systemic absorption ofthe polypeptide in the lower respiratory tract. The invention alsoprovides methods of manufacturing such formulations and the use of suchformulations in treating patients.

One approach for the pulmonary delivery of dry powder drugs utilizes ahand-held device with a hand pump for providing a source of pressurizedgas. The pressurized gas is abruptly released through a powderdispersion device, such as a venturi nozzle, and the dispersed powdermade available for patient inhalation.

Dry powder dispersion devices are described in several patents. U.S.Pat. No. 3,921,637 describes a manual pump with needles for piercingthrough a single capsule of powdered medicine. The use of multiplereceptacle disks or strips of medication is described in European PatentApplication No. EP 0 467 172; International Patent Publication Nos.WO91/02558; and WO93/09832; U.S. Pat. Nos. 4,627,432; 4,811,731;5,035,237; 5,048,514; 4,446,862; 5,048,514, and 4,446,862.

The aerosolization of protein therapeutic agents is disclosed inEuropean Patent Application No. EP 0 289 336. Therapeutic aerosolformulations are disclosed in International Patent Publication No. WO90/09781.

The present invention provides formulating Tf fusion protein for oralinhalation. The formulation comprises Tf fusion protein and suitablepharmaceutical excipients for pulmonary delivery. The present inventionalso provides administering the Tf fusion protein composition via oralinhalation to subjects in need thereof.

Transgenic Animals

The production of transgenic non-human animals that contain a modifiedtransferrin fusion construct with increased serum half-life increasedserum stability or increased bioavailability of the instant invention iscontemplated in one embodiment of the present invention. In someembodiments, lactoferrin may be used as the Tf portion of the fusionprotein so that the fusion protein is produced and secreted in milk.

The successful production of transgenic, non-human animals has beendescribed in a number of patents and publications, such as, for exampleU.S. Pat. No. 6,291,740 (issued Sep. 18, 2001); U.S. Pat. No. 6,281,408(issued Aug. 28, 2001); and U.S. Pat. No. 6,271,436 (issued Aug. 7,2001) the contents of which are hereby incorporated by reference intheir entireties.

The ability to alter the genetic make-up of animals, such asdomesticated mammals including cows, pigs, goats, horses, cattle, andsheep, allows a number of commercial applications. These applicationsinclude the production of animals which express large quantities ofexogenous proteins in an easily harvested form (e.g., expression intothe milk or blood), the production of animals with increased weightgain, feed efficiency, carcass composition, milk production or content,disease resistance and resistance to infection by specificmicroorganisms and the production of animals having enhanced growthrates or reproductive performance. Animals which contain exogenous DNAsequences in their genome are referred to as transgenic animals.

The most widely used method for the production of transgenic animals isthe microinjection of DNA into the pronuclei of fertilized embryos (Wallet al., J. Cell. Biochem. 49:113 [1992]). Other methods for theproduction of transgenic animals include the infection of embryos withretroviruses or with retroviral vectors. Infection of both pre- andpost-implantation mouse embryos with either wild-type or recombinantretroviruses has been reported (Janenich, Proc. Natl. Acad. Sci. USA73:1260 [1976]; Janenich et al., Cell 24:519 [1981]; Stuhlmann et al.,Proc. Natl. Acad. Sci. USA 81:7151 [1984]; Jahner et al., Proc. Natl.Acad Sci. USA 82:6927 [1985]; Van der Putten et al., Proc. Natl. AcadSci. USA 82:6148-6152 [1985]; Stewart et al., EMBO J. 6:383-388 [1987]).

An alternative means for infecting embryos with retroviruses is theinjection of virus or virus-producing cells into the blastocoele ofmouse embryos (Jahner, D. et al., Nature 298:623 [1982]). Theintroduction of transgenes into the germline of mice has been reportedusing intrauterine retroviral infection of the midgestation mouse embryoPalmer et al., supra [1982]). Infection of bovine and ovine embryos withretroviruses or retroviral vectors to create transgenic animals has beenreported. These protocols involve the micro-injection of retroviralparticles or growth arrested (i.e., mitomycin C-treated) cells whichshed retroviral particles into the perivitelline space of fertilizedeggs or early embryos (PCT International Application WO 90/08832 [1990];and Haskell and Bowen, Mol. Reprod. Dev., 40:386 [1995]. PCTInternational Application WO 90/08832 describes the injection ofwild-type feline leukemia virus B into the perivitelline space of sheepembryos at the 2 to 8 cell stage. Fetuses derived from injected embryoswere shown to contain multiple sites of integration.

U.S. Pat. No. 6,291,740 (issued Sep. 18, 2001) describes the productionof transgenic animals by the introduction of exogenous DNA intopre-maturation oocytes and mature, unfertilized oocytes (i.e.,pre-fertilization oocytes) using retroviral vectors which transducedividing cells (e.g., vectors derived from murine leukemia virus [MLV]).This patent also describes methods and compositions for cytomegaloviruspromoter-driven, as well as mouse mammary tumor LTR expression ofvarious recombinant proteins.

U.S. Pat. No. 6,281,408 (issued Aug. 28, 2001) describes methods forproducing transgenic animals using embryonic stem cells. Briefly, theembryonic stem cells are used in a mixed cell co-culture with a morulato generate transgenic animals. Foreign genetic material is introducedinto the embryonic stem cells prior to co-culturing by, for example,electroporation, microinjection or retroviral delivery. ES cellstransfected in this manner are selected for integrations of the gene viaa selection marker such as neomycin.

U.S. Pat. No. 6,271,436 (issued Aug. 7, 2001) describes the productionof transgenic animals using methods including isolation of primordialgerm cells, culturing these cells to produce primordial germcell-derived cell lines, transforming both the primordial germ cells andthe cultured cell lines, and using these transformed cells and celllines to generate transgenic animals. The efficiency at which transgenicanimals are generated is greatly increased, thereby allowing the use ofhomologous recombination in producing transgenic non-rodent animalspecies.

Gene Therapy

The use of modified transferrin fusion constructs for gene therapywherein a modified transferrin protein or transferrin domain is joinedto a therapeutic protein or peptide is contemplated in one embodiment ofthis invention. The modified transferrin fusion constructs withincreased serum half-life or serum stability of the instant inventionare ideally suited to gene therapy treatments.

The successful use of gene therapy to express a soluble fusion proteinhas been described. Briefly, gene therapy via injection of an adenovirusvector containing a gene encoding a soluble fusion protein consisting ofcytotoxic lymphocyte antigen 4 (CTLA4) and the Fc portion of humanimmunoglobulin GI was recently shown in Ijima et al. (Jun. 10,2001)-Human Gene Therapy (United States) 12/9:1063-77. In thisapplication of gene therapy, a murine model of type II collagen-inducedarthritis was successfully treated via intraarticular injection of thevector.

Gene therapy is also described in a number of U.S. patents includingU.S. Pat. No. 6,225,290 (issued May 1, 2001); U.S. Pat. No. 6,187,305(issued Feb. 13, 2001); and U.S. Pat. No. 6,140,111 (issued Oct. 31,2000).

U.S. Pat. No. 6,225,290 provides methods and constructs wherebyintestinal epithelial cells of a mammalian subject are geneticallyaltered to operatively incorporate a gene which expresses a proteinwhich has a desired therapeutic effect. Intestinal cell transformationis accomplished by administration of a formulation composed primarily ofnaked DNA, and the DNA may be administered orally. Oral or otherintragastrointestinal routes of administration provide a simple methodof administration, while the use of naked nucleic acid avoids thecomplications associated with use of viral vectors to accomplish genetherapy. The expressed protein is secreted directly into thegastrointestinal tract and/or blood stream to obtain therapeutic bloodlevels of the protein thereby treating the patient in need of theprotein. The transformed intestinal epithelial cells provide short orlong term therapeutic cures for diseases associated with a deficiency ina particular protein or which are amenable to treatment byoverexpression of a protein.

U.S. Pat. No. 6,187,305 provides methods of gene or DNA targeting incells of vertebrate, particularly mammalian, origin. Briefly, DNA isintroduced into primary or secondary cells of vertebrate origin throughhomologous recombination or targeting of the DNA, which is introducedinto genomic DNA of the primary or secondary cells at a preselectedsite.

U.S. Pat. No. 6,140,111 (issued Oct. 31, 2000) describes retroviral genetherapy vectors. The disclosed retroviral vectors include an insertionsite for genes of interest and are capable of expressing high levels ofthe protein derived from the genes of interest in a wide variety oftransfected cell types. Also disclosed are retroviral vectors lacking aselectable marker, thus rendering them suitable for human gene therapyin the treatment of a variety of disease states without theco-expression of a marker product, such as an antibiotic. Theseretroviral vectors are especially suited for use in certain packagingcell lines. The ability of retroviral vectors to insert into the genomeof mammalian cells has made them particularly promising candidates foruse in the genetic therapy of genetic diseases in humans and animals.Genetic therapy typically involves (1) adding new genetic material topatient cells in vivo, or (2) removing patient cells from the body,adding new genetic material to the cells and reintroducing them into thebody, i.e., in vitro gene therapy. Discussions of how to perform genetherapy in a variety of cells using retroviral vectors can be found, forexample, in U.S. Pat. Nos. 4,868,116, issued Sep. 19, 1989, and4,980,286, issued Dec. 25, 1990 (epithelial cells), WO89/07136 publishedAug. 10, 1989 (hepatocyte cells), EP 378,576 published Jul. 25, 1990(fibroblast cells), and WO89/05345 published Jun. 15, 1989 andWO/90/06997, published Jun. 28, 1990 (endothelial cells), thedisclosures of which are incorporated herein by reference.

Without further description, it is believed that a person of ordinaryskill in the art can, using the preceding description and the followingillustrative examples, make and utilize the present invention andpractice the claimed methods. For example, a skilled artisan wouldreadily be able to determine the biological activity, both in vitro andin vivo, for the fusion protein constructs of the present invention ascompared with the comparable activity of the therapeutic moiety in itsunfused state. Similarly, a person skilled in the art could readilydetermine the serum half life and serum stability of constructsaccording to the present invention. The following working examplestherefore, specifically point out the preferred embodiments of thepresent invention, and are not to be construed as limiting in any waythe remainder of the disclosure.

EXAMPLES Example 1 GLP-1-Transferrin Fusion Proteins

GLP-1 is a peptide that regulates insulin secretion. It possessesanti-diabetic activity in human subjects suffering diabetes, especiallytype II diabetes. Like other peptides, GLP-1 has a short plasmahalf-life in humans. The present invention provides fusion proteins withGLP-1 fused to mTf with increased half-life and pharmaceuticalcompositions of such fusion proteins for the treatment of diseasesassociated with abnormal glucose levels that can be administered orally.

The present invention also provides fusion proteins comprising an GLP-1analog and mTF. In one embodiment of the invention, the GLP-1 analogcomprises an additional His residue at the N-terminus. The His residuecould be added to the N-terminus of GLP-1 or inserted after the Hisresidue at the N-terminus of GLP-1. In another embodiment, the GLP-1analog comprises an amino acid substitution at position 2. For example,the Ala in GLP-1(7-36) or GLP-1(7-37) peptide (SEQ ID NO: 6) issubstituted with another amino acid.

In this example, the steps for producing a GLP-1/mTf fusion protein aredescribed. The same steps may be used to generate transferrin fusionproteins with analogs of the GLP-1 peptides.

To produce the GLP-1/mTf fusion protein, the amino acid sequence ofGLP-1(7-36) and GLP-1(7-37) may be used.

haegtftsdvssylegqaakefiawlvkgr (SEQ ID NO: 7)haegtftsdvssylegqaakefiawlvkgrg (SEQ ID NO: 8)For example, the peptide sequence of GLP-1(7-36) may be back translatedinto DNA and codon optimized for yeast:

catgctgaaggtacttttacttctgatgtttcttcttatttggaaggtcaagctgctaaagaa  h  a  e  g  t  f  t  s  d  v  s  s  y  l  e  g  q  a  a  k  etttattgcttggttggttaaaggtaga (SEQ ID NO: 13)  f  i  a  w  l  v  k  g  r (SEQ ID NO: 14)The primers were specifically designed to form 5′ XbaI and 3′ KpnIsticky ends after annealing and to enable direct ligation into XbaI/KpnIcut pREX0052, just 5′ of the end of the leader sequence and at theN-terminus of mTf. Alternatively, other sticky ends may be engineeredfor ligations into other vectors.

    ′XbaI    -+----- 1aggtctctag agaaaaggca tgctgaaggt acttttactt ctgatgtttc ttcttatttgtccagagatc tcttttccgt acgacttcca tgaaaatgaa gactacaaag aagaataaac >>......FL.......>>  r  s  l   e  k  r (SEQ ID NO: 17)                   >>..................GLP-1....................>                      h  a  e  g   t  f  t   s  d  v   s  s  y  l                                                     KpnI                                                   ------+ 61gaaggtcaag ctgctaaaga atttattgct tggttggtta aaggtagggt acctgatacttccagttc gacgatttct taaataacga accaaccaat ttccatccca tggactat >......................GLP-1......................>>  e  g  q   a  a  k   e  f  i  a   w  l  v   k  g  r                                                    >>..mTf..>>                                                       v  p  dTop strand: SEQ ID NO: 15 Bottom strand: SEQ ID NO:16Top strand primer: P0056 (nucleotides 7-112 of SEQ ID NO: 15)Bottom strand primer: P0057 (nucleotides 9-108 of SEQ ID NO: 16)

After annealing and ligation, the clones were sequenced to confirmcorrect insertion. This vector was designated pREX0094. The cassette wascut out of pREX0094 with NotI and sub-cloned into NotI cut yeast vector,pSAC35, to make pREX0100.

This plasmid was then electroporated into the host Saccharomyces yeaststrains and transformants selected for leucine prototrohy on minimalmedia plates. Expression was determined by growth in liquid minimalmedia and analysis of supernatant by SDS-PAGE, western blot, and ELISA.

Example 2 Insulin-Transferrin Fusion Proteins

Insulin is a peptide hormone that is secreted by the islets ofLangerhans in the pancreas and that regulates the metabolism ofcarbohydrates and fats, in particular the conversion of glucose toglycogen. It is given to humans suffering from type I and type IIdiabetes, as well as to diabetic animals. Currently, insulin must beadministered by subcutaneous injection and has a short plasma half-lifein humans. The present invention provides fusion proteins of insulinfused to mTf that have increased half-life and pharmaceuticalcompositions of such fusion proteins for the treatment of diseasesassociated with abnormal glucose levels that can be administered orally.

In this example, the steps for producing an insulin/mTf fusion proteinare described. Similar steps may be followed to generate transferrinfusion proteins with analogs of insulin peptides.

For expression in Saccharomyces constructs were initially made in thebase vector pREX0052, comprising an E. coli cloning vector with acassette for the expression of mTf in yeast, as either inserts betweenthe 5′ XbaI/KpnI sites for the N-terminal fusion, or 3′ SalI/HindIIIsites for the C-terminal fusion.

       XbaI         KpnI      -+-----      -------+aggtctctag agaagagggt acctgata (SEQ ID NO: 18)tccagagatc tcttctccca tggactat >>......FL.......>>   r  s  l   e  k  r (SEQ ID NO: 19)                    >>..mTf..>>                      v  p  d          SalI       HindIII       -+-----      --+---- actttccgtc gaccttaata agcttaattc(SEQ ID NO: 20) tgaaaggcag ctggaattat tcgaattaag >>........mTf........>>  t  f  r   r  p  -   - (SEQ ID NO: 21)                 mADHlt >>.....>>

These constructs form an expression cassette with (5′ to 3′) the yeastPRB1 promoter, leader sequence directing secretion into the growthmedia, (N-terminal fusion), mTf sequence, (C-terminal fusion), stopcodons and the ADH1 terminator sequence. Once constructed, theexpression cassettes were recovered as NotI fragments and inserted intoNotI digested pSAC35, an E. coli/yeast shuttle vector.

The insulin sequence used corresponds to that of Genbank Accession No.NM_(—)000207, SEQ ID NOS: 22 (DNA) and 23 (protein), as shown below.

1 gctgcatcag aagaggccat caagcacatc actgtccttc tgccatggcc ctgtggatgccgacgtagtc ttctccggta gttcgtgtag tgacaggaag acggtaccgg gacacctacg                                                >>.....INS......>                                                >>...leader.....>                                                  m  a   l  w  m 61gcctcctgcc cctgctggcg ctgctggccc tctggggacc tgacccagcc gcagcctttgcggaggacgg ggacgaccgc gacgaccggg agacccctgg actgggtcgg cgtcggaaac >..............................INS..............................> >..........................leader..........................>> r  l  l   p  l  l  a   l  l  a   l  w  g   p  d  p  a   a  a                                                            >>.>                                                              f 121tgaaccaaca cctgtgcggc tcacacctgg tggaagctct ctacctagtg tgcggggaacacttggttgt ggacacgccg agtgtggacc accttcgaga gatggatcac acgccccttg >..............................INS..............................> >...............................B...............................> v  n  q   h  l  c  g   s  h  l   v  e  a   l  y  l  v   c  g  e 181gaggcttctt ctacacaccc aagacccgcc gggaggcaga ggacctgcag gtggggcaggctccgaagaa gatgtgtggg ttctgggcgg ccctccgtct cctggacgtc caccccgtcc >..............................INS..............................>                              r   r  e  a   e  d  l  q   v  g  q >............B............>> r  g  f   f  y  t  p   k  t 241tggagctggg cgggggccct ggtgcaggca gcctgcagcc cttggccctg gaggggtcccacctcgaccc gcccccggga ccacgtccgt cggacgtcgg gaaccgggac ctccccaggg >..............................INS..............................> v  e  l   g  g  g  p   g  a  g   s  l  q   p  l  a  l   e  g  s 301tgcagaagcg tggcattgtg gaacaatgct gtaccagcat ctgctccctc taccagctggacgtcttcgc accgtaacac cttgttacga catggtcgta gacgagggag atggtcgacc >..............................INS..............................> l  q  k   r            >>........................A.........................>              g  i  v   e  q  c   c  t  s   i  c  s  l   y  q  l 361agaactactg caactagacg cagcccgcag gcagcccccc acccgccgcc tcctgcaccgtcttgatgac gttgatctgc gtcgggcgtc cgtcgggggg tgggcggcgg aggacgtggc >......INS......>> >......A.....>> e  n  y   c  n 421 agagagatgg aataaagccc ttgaaccagctctctctacc ttatttcggg aacttggtcg

The cDNA for the above sequence can be generated in a number of ways,e.g., by RT-PCR, from a cDNA pool, or by overlapping syntheticoligonucleotides. To generate a clone from a cDNA pool, two primers weresynthesized and used as PCR primers.

5′ primer (SEQ ID NO: 24) 5′-tttgtgaaccaacacctgtgcggc-3′ 3′ primer(SEQ ID NO: 25) 3′-gacgagggagatggtcgacctcttgatgacgttg-5′

To make the N-terminal insert, a 5′ mutagenic primer was used to createa second PCR product using the first PCR product as template. Thisprimer inserted the last 5 amino acids of the leader sequence and theXbaI site. The KpnI site could not be inserted by this method, as anamino acid change would have resulted from the creation of the KpnIsite. Instead, the PCR product was digested with XbaI/PvuII. A linkerwas then made of two overlapping oligos with a PvuII 5′ end and 3′overhang which would ligate to the KpnI overhang on KpnI digestedpREX0052. By annealing and ligating this linker to the digested PCRfragment and ligating the resulting product into XbaI/KpnI digestedpREX0052 the plasmid pREX0052 N-insulin (SEQ ID NOS: 26 and 27) wasgenerated.

                XbaI                -+---- 1gcttactcta ggtctctaga taagaggttt gtgaaccaac acctgtgcgg ctcacacctgcgaatgagat ccagagatct attctccaaa cacttggttg tggacacgcc gagtgtggac >>...........FL............>>  a  y  s   r  s  l  d  k   r                             >>..............Ins................>                             >>...............B.................>                               f   v  n  q   h  l  c   g  s  h  l 61gtggaagctc tctacctagt gtgcggggaa cgaggcttct tctacacacc caagacccgccaccttcgag agatggatca cacgcccctt gctccgaaga agatgtgtgg gttctgggcg >..............................Ins..............................>                                                                r >.............................B.............................>>  v  e  a   l  y  l   v  c  g  e   f  g  f   f  y  t   p  k  t 121cgggaggcag aggacctgca ggtggggcag gtggagctgg gcgggggccc tggtgcaggcgccctccgtc tcctggacgt ccaccccgtc cacctcgacc cgcccccggg accacgtccg >..............................Ins..............................>  r  e  a   e  d  l   q  v  g  q   v  e  l   g  g  g   p  g  a  g 181agcctgcagc ccttggccct ggaggggtcc ctgcagaagc gtggcattgt ggaacaatgctcggacgtcg ggaaccggga cctccccagg gacgtcttcg caccgtaaca ccttgttacg >..............................Ins..............................>  s  l  q   p  l  a   l  e  g  s   l  q  k   r                                              >>.......A........>                                                 g  i  v  e  q c                          PvuII                           --+--- 241tgtaccagca tctgctccct ctaccagctg gagaactact gcaacgtacacatggtcgt agacgaggga gatggtcgac ctcttgatga cgttg >......................Ins.....................>> >.......................A......................>>  c  t  s   i  c  s   l  y  q  l   e  n  y   c  n                                                 >>mTf                                                   v 5′primer: 5′-gcttactctaggtctctagataagaggtttgtgaaccaacacctgtgcg-3′(SEQ ID NO: 28) Linkers: 5'-ctggagaactactgcaacgtac-3′ (SEQ ID NO: 29)3′-gacctcttgatgacgttg-5′ (SEQ ID NO: 30)

To make the C terminal insert, 5′ and 3′ mutagenic primers were used tocreate a second PCR product using the first PCR product as template.This product was then digested with SalI/HindIII and ligated intoSalI/HindIII digested pREX0052. This resulted in the plasmid pREX0052C-insulin (SEQ ID NOS: 31 and 32).

           SalI            -+---- 1tgcactttcc gtcgaccttt tgtgaaccaa cacctgtgcg gctcacacct ggtggaagctacgtgaaagg cagctggaaa acacttggtt gtggacacgc cgagtgtgga ccaccttcga >>......mTf......>>  c  t  f   r  r  p                   >>...................Ins.....................>                   >>....................B......................>                      f  v  n  q   h  l  c   g  s  h   l  v  e  a 61ctctacctag tgtgcgggga acgaggcttc ttctacacac ccaagacccg ccgggaggcagagatggatc acacgcccct tgctccgaag aagatgtgtg.ggttctgggc ggccctccgt >..............................Ins..............................>                                                       r  r  e  a >........................B........................>>  l  y  l   v  c  g   e  r  g  f   f  y  t   p  k  t 121gaggacctgc aggtggggca ggtggagctg ggcgggggcc ctggtgcagg cagcctgcagctcctggacg tccaccccgt ccacctcgac ccgcccccgg gaccacgtcc gtcggacgtc >.............................Ins...............................>  e  d  l   q  v  g   q  v  e  l   g  g  g   p  g  a   g  s  l  q 181cccttggccc tggaggggtc cctgcagaag cgtggcattg tggaacaatg ctgtaccagcgggaaccggg acctccccag ggacgtcttc gcaccgtaac accttgttac gacatggtcg >..............................Ins..............................>  p  l  a   l  e  g   s  l  q  k   r                                    >>............A.............>                                      g  i   v  e  q   c  c  t  s                                            HindIII                                            -+---- 241atctgctccc tctaccagct ggagaactac tgcaactaat aagcttaatttagacgaggg agatggtcga cctcttgatg acgttgatta ttcgaattaa >.................Ins................>> >..................A.................>>  i  c  s   l  y  q   l  e  n  y   c  n                                       mADH1t >>....>>                                                a  - 5′primer: 5′-tgcactttccgtcgaccttttgtgaaccaacacctgtgcg-3′ (SEQ ID NO: 33)3′ primer: 3′-gacctcttgatgacgttgattattcgaattaa-5′ (SEQ ID NO: 34)

Once the DNA sequence for both the N- and C-terminal inserts had beenchecked and confirmed, the plasmids pREX0052 N-insulin and pREX0052C-insulin were digested with NotI and the expression cassettesrecovered. These were then ligated into NotI digested pSAC35 to givepSAC35 N-insulin and pSAC35 C-insulin. These plasmids were thenelectroporated into the host Saccharomyces yeast strains andtransformants selected for leucine prototrohy on minimal media plates.Expression was determined by growth in liquid minimal media and analysisof superanatant by SDS-PAGE, western blot, ELISA and BIAcore.

These fusion constructs result in the production of proinsulin attachedto transferrin. Proteases in yeast may convert the proinsulin to insulinas it is being made and secreted, although the final expression productmay contain only proinsulin. In that case, the proinsulin can beconverted to insulin post-expression using an appropriate purifiedprotease.

Oral Administration of Insulin/Modified Transferrin Fusion Protein toRats

To test the insulin activity of the insulin/mTf fusion protein, diabeticrats are first prepared. Female Sprague-Dawley rats are fasted for 24hours and their blood glucose level determined to establish a baseline.The rats are then injected intraperitoneally with a solution ofstreptozotocin (STZ), 60 mg/ml, at a dosage of 60 mg/kg. I.p. injectionsof STZ are continued for four more days, and rats with a fasting bloodglucose level above 300 mg/dl are selected as diabetic rats.

Solutions of the fusion protein and of insulin alone are prepared in PBSor sodium bicarbonate to provide dosages of 7 to 80 units of insulin/kgwhen administered to rats. As a control, rats are also treated with PBSalone. The solutions or PBS are administered by oral gavage to ratsfollowing a 12 hour fast, and blood samples are collected from the tailafter 0, 30 and 60 minutes, and then at 2-hour intervals. Blood glucoselevels at 0, 0.5, 1, 3, 5, 7, 9 and 11 hours after dosing are measuredwith a blood glucose monitoring device designed for diabetics, and therats are fed again at 11 hours post-dose.

The activity of the insulin is determined by measuring the decrease inblood glucose level over time, correcting the decrease by any increasesor decreases in the PBS-only samples. The insulin activities of thefusion protein versus unfused insulin are compared.

To examine the uptake of the fusion protein by transferrin receptors inthe intestinal mucosa, fusion protein and unfused insulin as a controlare administered to diabetes induced rats as described above andtransport measured using standard sandwich ELISAs and serum samples.Alternatively, ¹²⁵I-labeled fusion protein or ¹²⁵I-labeled unfusedinsulin is adminstered to diabetes-induced rats at dosage of 80 Uinsulin/kg by oral gavage, as described above. Blood samples arecollected from the tail after 0, 30 and 60 minutes, then at 2-hourintervals, also as described above, and serum samples are analyzed byHPLC, using, for example a Sephacryl column and eluting samples withPBS. Standards containing ¹²⁵I-labeled transferrin, ¹²⁵I-labeled insulinand ¹²⁵I-labeled fusion protein are also run on the Sephacryl column todetermine their peak elution times and fraction numbers. Theradioactivity of each fraction is measured with a gamma counter, and theprotein content of each fraction is measured by the absorbance at 280nm. Serum samples from rats treated with the fusion protein may not showthe appearance of the fusion protein immediately, as there may be adelay of a few hours.

Example 3 Preparation of Therapeutic mTF Fusion Proteins with IncreasedIron Affinity

Therapeutic mTf fusion protein with increased iron affinity may beprepared. As an example for preparing modified transferrin fusionproteins with increased iron binding ability, the procedure in Example 2above may be carried out with the following modification. These fusionproteins may be used to facilitate uptake and transfer of the fusionprotein across the gastrointestinal epithelium.

A cloning vector such as pREX0052, described above, which contains themTf sequence is cut with a restriction enzyme, or a pair of restrictionenzymes, to remove a portion of the mTf gene. Using techniques standardin the art, this fragment is then subjected to site-directed mutagenesisusing primers that introduce a mutation at a position corresponding tonucleotide 723 of SEQ ID NO: 1, converting the codon AAG (Lys) to CAG(Gln) or GAG (Glu). Similarly, primers are used that introduce mutationsat positions corresponding to nucleotides 726 and 728 of SEQ ID NO: 1,converting the codon CAC (His) to CAG (Gln) or GAG (Glu). Primers mayalso be used that introduce mutations at all three nucleotide positions,resulting in the substitution of two adjacent amino acids. Thesenucleotide positions correspond to amino acids 225 and 226 of theprotein encoded with the leader sequence and to amino acids 206 and 207of the mature protein. The mutated fragment is then amplified by RT-PCRand religated into the cloning vector. This vector containing themutation or mutations is used in a subsequent step for introduction of aDNA molecule coding for the therapeutic protein. As described in Example2, above, the mTf fusion protein sequence may be introduced into yeastexpression vectors and transformed into Saccharomyces or other yeastsfor protein production.

As discussed previously, other amino acids may also be mutated to obtaintherapeutic mTf proteins with increased iron affinity.

Example 4 β-Interferon-Transferrin Fusion Proteins

β-IFN is effective in the treatment of various diseases, such as, butnot limited to, multiple sclerosis, brain tumor, skin cancer, andhepatitis B and C. Like most cytokines, β-IFN has a short circulationhalf-life. The present invention provides fusion proteins comprisingβ-IFN fused to mTf with increased half-life and efficacy in patients.This example describes the steps in generating a β-IFN/mTf fusionprotein that may be administered orally.

In this example, IFNβ-1 is fused to modified transferrin at either theN- or C-termini. The IFNβ-1 clone was obtained from the ATCC (No.39517). Specifically designed primers were used to confirm the DNAsequence of the IFNβ-1 clone. These primers were external to the IFNβ-1DNA sequence and designed to read in from the vector such that thefull-length sequence of the clone was obtained. The primers used were:

P0070 GCTATGACCAACAAGTGTCTC35) (SEQ ID NO: 35) andP0071 CGCACCTGTGGCGCCGGTGATG (SEQ ID NO: 36)

N-Terminal Fusion

Once the DNA sequence was confirmed, primers were designed for fusion ofIFNβ-1 to mTf. The N-terminal fusion was a two step process. A straightfusion using primers with XbaI and KpnI sites would have destroyed theKpnI site and clipped the beginning of mTf. A linker, primers P0082(nucleotides 18-48 of SEQ ID NO: 37) and P0083 (nucleotides 17-39 SEQ IDNO: 38), was designed to create an internal KpnI site at the 3′ end ofIFNβ-1, by a single silent mutation of by 486 from T to G (bold), andwith a 5′ XbaI overhang and 3′ GTAC which would anneal with a KpnI site.The overhang destroyed the existing KpnI site in pREX0052. The linkerswere annealed and ligated into pREX0054 cut with XbaI/KpnI, creating anintermediate vector with mTf untouched and a KpnI site that could beused to fuse the IFNβ-1 gene at the N-terminus of mTf.

                   XbaI           KpnI                 -+-----         -----+                  >>............P0082.............>>ctgcttactc taggtctcta gagaaaacag ggtacctccg aaacgtacct gataaaactggacgaatgag atccagagat ctcttttgtc ccatggaggc tttgcatgga ctattttgac                       <<........P0083........<< >>.........MFa-1..........>>    a  y   s  r  s  l   e  k (SEQ ID NO: 39)                            >>....IFN-B-1.....>>                              t   g  y  l   r  n (SEQ ID NO: 40)                                                >>.....mTf......>                                                  v  p   d  k  t                                                  (SEQ ID NO: 41)Top Strand: SEQ ID NO: 37 Bottom Strand: SEQ ID NO: 38

A second set of primers, P0084 (SEQ ID NO: 42) and P0085 (SEQ ID NO:43), were designed to tailor the ends of the IFNβ-1 gene by mutagenicPCR for subsequent insertion into the intermediate vector via the XbaIand KpnI sites. A XbaI/KpnI digest of this tailored gene removed thelast 5 amino acids of IFNβ-1; however, these were already engineeredinto the intermediate vector. The resulting construct, pREX0048, wascreated by ligating the IFNβ-1 gene cut with XbaI/KpnI into theXbaI/KpnI cut intermediate vector.

P0084 (SEQ ID NO: 42) >>-------FL--------->>                      >>----IFNβ-l------->>                            XbaI CTCTAGGTC

GAAAAGGAGCTACAACTTGCTTGGATTC P0085 (SEQ ID NO: 43)<<--------IFNβ-l-------<<           KpnI GTTTCGGA

CTGTAAGTCTG

After the pREX0048 construct was created, it was sequenced to confirmcorrect insertion. The expression cassette, as a NotI fragment, was thensub-cloned into NotI cut yeast vector, pSAC35, to make the pREX0050.

C-Terminal Fusion

Specifically designed primers, P0086 (SEQ ID NO: 44) and P0087 (SEQ IDNO: 45), were used to PCR amplify the original clone and, in addition,to tailor the ends of IFNβ-1 to have SalI and HindIII sites at the 5′and 3′ ends, respectively. The newly tailored product was ligated intoSalI/HindIII cut pREX0052 to create pREX0049.

P0086 (SEQ ID NO.: 44) >>---mTf---->>              >>-----IFNβ-l------->>          SalI ACTTTCC

CTAGCTACAACTTGCTTGGATTC P0087 (SEQ ID NO: 45) <<----3′ADHlt-------<<*  *                            <<-----IFNβ-l-------<<                 HindIII CATAAATCATAAGAATT

TATTAGTTTCGGAGGTAACCTGTAAGT

After the Prex0049 construct was created, it was sequenced to confirmcorrect insertion. The expression cassette, as a NotI fragment, is thensub-cloned into NotI cut yeast vector, such as pSAC35, to make pREX0051.

In one embodiment of the invention, β-IFN-1 (GenBank Acc. No.NM_(—)002176, SEQ ID NO: 46) is made more stable and soluble by mutatingCys17 (in the mature protein) to Ser. The mutation of Cys17 to Ser canbe performed by routine mutagenic reactions such as a mutagenic PCRreaction using specifically designed primers and the nucleic acidencoding β-IFN-1 as the template.

Further, the β-IFN-1 is modified to prevent glycosylation by modifyingthe N-linked glycosylation site, NES/T (residues 80 to 82 of SEQ ID NO:46). As an example, N could be mutagenized to Q and S/T could bemutagenized to Ala or other amino acid acids. Such mutagenesis could beperformed with mutagenic PCR reaction using specifically designedprimers and the nucleic acid encoding β-IFN-1 as the template.

Although the present invention has been described in detail withreference to examples above, it is understood that various modificationscan be made without departing from the spirit of the invention.Accordingly, the invention is limited only by the following claims. Allcited patents, patent applications and publications referred to in thisapplication are herein incorporated by reference in their entirety.

1. A pharmaceutical composition formulated for oral delivery, nasaldelivery, or pulmonary delivery comprising a transferrin (Tf) proteinexhibiting reduced glycosylation fused to at least one therapeuticprotein or peptide.
 2. A pharmaceutical composition of claim 1,formulated to deliver the Tf fusion protein to the gastrointestinalepithelium of a patient.
 3. A pharmaceutical composition of claim 2,wherein the Tf protein binds to the transferrin receptor on thegastrointestinal epithelium when orally administered to a patient.
 4. Apharmaceutical compositions of claim 2, wherein iron is bound to the Tfprotein
 5. A pharmaceutical formulation of claim 2, wherein the Tfprotein has been modified to increase its affinity or avidity for one ormore of the following selected from group consisting of transferrinreceptor, iron ions, and carbonate ions.
 6. A pharmaceutical compositionof claim 1, wherein the serum half-life of the therapeutic protein orpeptide is increased over the serum half-life of the therapeutic proteinor peptide in an unfused state.
 7. A pharmaceutical composition of claim1, wherein the therapeutic protein or peptide is fused to the C-terminalend of Tf.
 8. A pharmaceutical composition of claim 1, wherein thetherapeutic protein or peptide is fused to the N-terminal end of Tf. 9.A pharmaceutical composition of claim 1, wherein the therapeutic proteinor peptide is inserted into at least one loop of the Tf.
 10. Apharmaceutical composition of claim 1, wherein the Tf protein is lactotransferrin (lactoferrin) or a melanotransferrin.
 11. A pharmaceuticalcomposition of claim 1, wherein said Tf protein comprises at least onemutation that reduces or prevents glycosylation.
 12. A pharmaceuticalcomposition of claim 1, wherein the Tf fusion protein is expressed inthe presence of a compound that inhibits glycosylation.
 13. Apharmaceutical composition of claim 1, wherein said Tf fusion proteincomprises N terminal to C terminal: a therapeutic protein or peptide, alinker and Tf.
 14. A pharmaceutical composition of claim 13, wherein thelinker is a peptide that links the therapeutic protein or peptide to Tf15. A pharmaceutical composition of claim 1, wherein said therapeuticprotein or peptide is inserted between an N and a C domain of Tf.
 16. Apharmaceutical composition of claim 1, wherein the Tf protein has atleast one amino acid substitution, deletion or addition.
 17. Apharmaceutical composition of claim 1, wherein said Tf protein comprisesa first portion of the N domain of a Tf protein, a bridging peptide anda second portion of the N domain of a Tf protein, wherein the first andsecond portions are the same.
 18. A pharmaceutical composition of claim5, wherein said Tf protein has at least one amino acid substitution,deletion or addition at a position selected from the group consisting ofLys 206, His207 and a combination thereof.
 19. A pharmaceuticalcomposition of claim 1, wherein the formulation comprises iron bound tothe transferrin.
 20. A pharmaceutical composition of claim 1, whereinthe iron is bound to the fusion protein.
 21. A pharmaceuticalcomposition of claim 11, wherein the glycosylation site is selected fromthe group consisting of an amino acid residue corresponding to aminoacids N413 or N611.
 22. A pharmaceutical composition of claim 1, whereinthe fusion protein is effective to treat a human disease.
 23. Apharmaceutical composition of claim 22, wherein the fusion protein iseffective to treat a chronic human disease.
 24. A pharmaceuticalcomposition of claim 23, wherein the chronic disease is selected fromthe group consisting of a viral disease, cancer, a metabolic disease,obesity, an autoimmune disease, an inflammatory disease, an allergy,graft-vs.-host disease, systemic microbial infection, anemia,cardiovascular disease, a neurodegenerative disease, a disorder ofhematopoietic cells, diseases of the endocrine system or reproductivesystems, gastrointestinal diseases, diabetes and multiple sclerosis. 25.A pharmaceutical composition of claim 24, wherein the therapeuticprotein or peptide is selected from the group consisting of insulin,proinsulin, an insulin analog or derivative, GLP-1 and a GLP-1 analog orderivative.
 26. A pharmaceutical composition of claim 1, wherein thecomposition is formulated with an enteric coating.
 27. A pharmaceuticalcomposition of claim 1, wherein the fusion protein is dispersed in apharmaceutically acceptable carrier.
 28. A pharmaceutical composition ofclaim 27, wherein the pharmaceutically acceptable carrier is selectedfrom the group consisting of aqueous buffers, sucrose, lactose, starch,fatty oils, fatty acid esters, polysaccharides, monoglycerides,triglycerides, phospholipid emulsifiers, non-ionic emulsifiers andrefined colloid clays.
 29. A pharmaceutical composition of claim 1,wherein the fusion protein is contained in a solid form and wherein thesolid form is selected from the group consisting of a tablet, a chewabletablet, a capsule, a granulate or a powder.
 30. A pharmaceuticalcomposition of claim 29, wherein the tablet or capsule isenteric-coated.
 31. A pharmaceutical composition of claim 29, whereinthe capsule is a soft gelatin capsule.
 32. A pharmaceutical compositionof claim 29, wherein the solid form is formulated for slow release inthe gut.
 33. A pharmaceutical composition of claim 1, wherein the fusionprotein is formulated as a liquid, aerosol or syrup.
 34. Apharmaceutical composition of claim 1, formulated to comprise betweenabout 1 pg/kg and about 100 mg/kg body weight of the fusion protein. 35.A pharmaceutical composition of claim 1, formulated to comprise betweenabout 100 ng/kg and about 100 μg/kg body weight of the fusion protein.36. A pharmaceutical composition of claim 1, formulated to comprisebetween about 100 μg/kg and about 100 mg/kg body weight of the fusionprotein.
 37. A pharmaceutical composition of claim 18, wherein thetransferrin comprises bound iron.
 38. A pharmaceutical composition ofclaim 18, wherein the substituted amino acid is glutamine or glutamicacid.
 39. A pharmaceutical composition of claim 38, wherein the lysineresidue at amino acid 206 is replaced with a glutamine and the histidineresidue at amino acid 207 is replaced with a glutamic acid.
 40. Apharmaceutical composition of claim 1, wherein the pharmaceuticalcomposition is formulated to be mixed with food or a beverage.
 41. Apharmaceutical composition of claim 1, wherein the pharmaceuticalcomposition is formulated as a feed supplement for veterinary use.
 42. Apharmaceutical composition of claim 1, comprising between about 1 μg and1 g of fusion protein.
 43. A pharmaceutical composition of claim 42,comprising between about 10 μg and 100 mg of fusion protein.
 44. Apharmaceutical composition of claim 42, comprising between about 10 mgand 50 mg of fusion protein.
 45. A pharmaceutical composition formulatedfor oral delivery comprising a transferrin (Tf) protein exhibitingreduced glycosylation fused to at least one insulin protein or peptide.46. A pharmaceutical composition of claim 45, wherein the insulinprotein or peptide is human insulin.
 47. pharmaceutical composition ofclaim 46, wherein human insulin comprises SEQ ID NO:
 19. 48. Apharmaceutical composition of claim 45, wherein the insulin protein orpeptide is fused to the N-terminal end of transferrin.
 49. Apharmaceutical composition of claim 48, wherein the insulin protein orpeptide is separated from the N-terminal end of transferrin by a linkerpeptide.
 50. A pharmaceutical composition of claim 45, wherein thetransferrin comprises iron or carbonate ions.
 51. A pharmaceuticalcomposition of claim 45, wherein the insulin protein or peptide isproinsulin.
 52. A pharmaceutical composition of claim 45, wherein theinsulin protein or peptide is mature human insulin.
 53. A pharmaceuticalcomposition of claim 45, formulated to comprise between about 5 to about500 units of insulin per kg of patient weight.
 54. A pharmaceuticalcomposition of claim 53, formulated to comprise between about 10 toabout 100 units of insulin per kg of patient weight.
 55. Apharmaceutical composition of claim 45, formulated to comprise aneffective amount of insulin to induce a decrease in blood glucose in apatient.
 56. A pharmaceutical composition of claim 55, formulated tocomprise an effective amount of insulin to induce a hypoglycemic effectin a patient.
 57. A pharmaceutical composition of claim 55, wherein theeffective amount decreases blood glucose levels by about 5% to about80%.
 58. A pharmaceutical composition of claim 45, formulated toneutralize or protect the fusion protein from gastric acid and/orenzymes.
 59. A pharmaceutical composition of claim 45, furthercomprising a transcytosis enhancer.
 60. A pharmaceutical composition ofclaim 45, formulated to comprise an effective amount of fusion proteinto increase the serum insulin activity level in a patient.
 61. Apharmaceutical composition of claim 60, wherein the patient is a human.62. A pharmaceutical composition of claim 61, wherein the patient is ajuvenile or adult.
 63. A pharmaceutical composition of claim 62, whereinthe patient is a diabetic patient.
 64. A pharmaceutical composition ofclaim 63, wherein the patient has juvenile- or adult-onset diabetes. 65.A pharmaceutical composition of claim 64, wherein the patient has TypeII diabetes.
 66. A pharmaceutical composition formulated for oraldelivery comprising a transferrin (Tf) protein exhibiting reducedglycosylation fused to at least one GLP-1 protein or peptide.
 67. Apharmaceutical composition of claim 66, wherein the GLP-1 protein orpeptide is human GLP-1 amino acids 7-35, 7-36 or 7-37.
 68. Apharmaceutical composition of claim 66, wherein the human GLP-1 is GLP-1amino acids 7-37
 69. A pharmaceutical composition of claim 66, whereinthe GLP-1 protein or peptide is fused to the N-terminal end oftransferrin.
 70. A pharmaceutical composition of claim 69, wherein theGLP-1 protein or peptide is separated from the N-terminal end oftransferrin by a linker peptide.
 71. A pharmaceutical composition ofclaim 66, wherein the transferrin comprises iron or carbonate ions. 72.A pharmaceutical composition of claim 66, wherein the GLP-1 protein orpeptide has a modified N-terminal end to prevent cleavage.
 73. Apharmaceutical composition of claim 66, wherein the modification is atleast one amino acid substitution.
 74. A pharmaceutical composition ofclaim 66, formulated to comprise an effective amount of GLP-1 to inducea decrease in blood glucose in a patient.
 75. A pharmaceuticalcomposition of claim 66, formulated to comprise an effective amount ofGLP-1 to induce a hypoglycemic effect in a patient.
 76. A pharmaceuticalcomposition of claim 74, wherein the effective amount decreases bloodglucose levels by about 5% to about 80%.
 77. A pharmaceuticalcomposition of claim 66, formulated to neutralize or protect the fusionprotein from gastric acid and/or enzymes.
 78. A pharmaceuticalcomposition of claim 66, further comprising a transcytosis enhancer. 79.A pharmaceutical composition of claim 66, formulated to comprise aneffective amount of fusion protein to increase the serum GLP-1 activitylevel in a patient.
 80. A pharmaceutical composition of claim 79,wherein the patient is a human.
 81. A pharmaceutical composition ofclaim 80, wherein the patient is a juvenile or adult.
 82. Apharmaceutical composition of claim 81, wherein the patient is adiabetic patient.
 83. A pharmaceutical composition of claim 82, whereinthe patient has juvenile- or adult-onset diabetes.
 84. A pharmaceuticalcomposition of claim 83, wherein the patient has Type II diabetes.
 85. Amethod of administering a therapeutic protein or peptide to a patient inneed thereof, comprising orally administering a pharmaceuticalcomposition of claim
 1. 86. A method of enhancing the absorption of atherapeutic protein or peptide into the bloodstream from the digestivetract of a patient in need thereof, comprising: a) administering afusion protein of claim 1, wherein the therapeutic protein or peptide isabsorbed into the bloodstream at an increased rate compared toabsorption of the therapeutic protein or peptide in an unfused state.87. A method of claim 85, further comprising administering atranscytosis enhancer.
 88. A method of claim 86, further comprisingadministering a transcytosis enhancer.
 89. A method of treating diabetesin a patient in need thereof, comprising orally administering a fusionprotein comprising a transferrin (Tf) protein exhibiting reducedglycosylation fused to at least one insulin protein or peptide.
 90. Amethod of treating diabetes in a patient in need thereof, comprisingorally administering a fusion protein comprising a transferrin (Tf)protein exhibiting reduced glycosylation fused to at least one GLP-1protein or peptide.
 91. A method of enhancing the absorption of aninsulin protein or peptide into the bloodstream from the digestive tractof a diabetic patient, comprising: a) administering a fusion proteincomprising a transferrin (Tf) protein exhibiting reduced glycosylationfused to at least one insulin protein or peptide.
 92. A method ofenhancing the absorption of a GLP-1 protein or peptide into thebloodstream from the digestive tract of a diabetic patient, comprising:a) administering a fusion protein comprising a transferrin (Tf) proteinexhibiting reduced glycosylation fused to at least one GLP-1 protein orpeptide.
 93. A method of altering the blood glucose levels in a patientin need thereof, comprising: a) administering a fusion proteincomprising a transferrin (Tf) protein exhibiting reduced glycosylationfused to at least one insulin protein or peptide.
 94. A method ofaltering the blood glucose levels in a patient in need thereof,comprising: a) administering a fusion protein comprising a transferrin(Tf) protein exhibiting reduced glycosylation fused to at least oneGLP-1 protein or peptide.
 95. The pharmaceutical composition of claim12, wherein the compound is tunicamycin.
 96. A pharmaceuticalcomposition of claim 1, wherein the formulation comprises carbonatebound to transferrin.
 97. A pharmaceutical composition of claim 1,wherein the formulation comprises and iron and carbonate bound totransferrin.
 98. A pharmaceutical composition of claim 11, wherein themutation is within the N-X-S/T glycosylation site.
 99. A pharmaceuticalcomposition of claim 98, wherein Ser or Thr is mutated.
 100. Apharmaceutical composition of claim 98, wherein X is mutated to Pro.101. A pharmaceutical composition of claim 25, wherein the GLP-1 analogis exendin.
 102. A pharmaceutical composition of claim 66, wherein theGLP-1 protein or peptide comprises the second residue from theN-terminus is substituted with another amino acid.
 103. A method ofclaim 93 or 94, wherein the patient is diabetic or obese.
 104. Apharmaceutical composition of claim 1, wherein the Tf protein comprisesa single N domain.